1-aryl-4-substituted isoquinolines

ABSTRACT

1-aryl-4-substituted isoquinoline or 1-aryl-3,4-disubstituted isoquinoline analogues of Formula I and Formula II, as follows: 
                         
wherein R 1 , R 2 , R 3 , R 8 , R 9 , A and Ar are defined herein. Such compounds are ligands of C5a receptors. Preferred compounds of Formula I and II bind to C5a receptors with high affinity and exhibit neutral antagonist or inverse agonist activity at C5a receptors. The present invention also relates to pharmaceutical compositions comprising such compounds, and to the use of such compounds in treating a variety of inflammatory, cardiovascular, and immune system disorders. In addition, the present invention provides labeled 1-aryl-4-substituted isoquinolines or 1-aryl-3,4-disubstituted isoquinolines, which are useful as probes for the localization of C5a receptors.

FIELD OF THE INVENTION

This invention relates generally to 1-aryl-4-substituted isoquinolinesthat act as modulators of mammalian complement C5a receptors, and topharmaceutical compositions comprising such modulators. The presentinvention further relates to the use of such modulators in treating avariety of inflammatory and immune system disorders and as probes forthe localization of C5a receptors.

BACKGROUND OF THE INVENTION

C5a, a 74 amino acid peptide, is generated in the complement cascade bythe cleavage of the complement protein C5 by the complement C5convertase enzyme. C5a has both anaphylatoxic (e.g., bronchoconstrictingand vascular spasmogenic) and chemotactic effects. Therefore, it isactive in engendering both the vascular and cellular phases ofinflammatory responses. Because it is a plasma protein and, therefore,generally almost instantly available at a site of an inciting stimulus,it is a key mediator in terms of initiating the complex series of eventsthat results in augmentation and amplification of an initialinflammatory stimulus. The anaphylatoxic and chemotactic effects of theC5a peptide are believed to be mediated through its interaction with theC5a receptor (CD88 antigen), a 52 kD membrane bound G-protein coupledreceptor (GPCR). C5a is a potent chemoattractant for polymorphonuclearleukocytes, bringing neutrophils, basophils, eosinophils and monocytesto sites of inflammation and/or cellular injury. C5a is one of the mostpotent chemotactic agents known for a wide variety of inflammatory celltypes. C5a also “primes” or prepares neutrophils for variousantibacterial functions (e.g., phagocytosis). Additionally, C5astimulates the release of inflammatory mediators (e.g., histamines,TNF-α, IL-1, IL-6, IL-8, prostaglandins, and leukotrienes) and therelease of lysosomal enzymes and other cytotoxic components fromgranulocytes. Among its other actions, C5a also promotes the productionof activated oxygen radicals and the contraction of smooth muscle.

Considerable experimental evidence implicates increased levels of C5a ina number of autoimmune diseases and inflammatory and related disorders.Agents that block the binding of C5a to its receptor other agents,including inverse agonists, which modulate signal transductionassociated with C5a-receptor interactions, can inhibit the pathogenicevents, including chemotaxis, associated with anaphylatoxin activitycontributing to such inflammatory and autoimmune conditions. The presentinvention provides such agents, and has further related advantages.

SUMMARY OF THE INVENTION

The present invention provides compounds that modulate, and preferablyinhibit, C5a receptor activation and/or C5a receptor-mediated signaltransduction. Such C5a receptor modulators are preferably high affinityC5a receptor ligands and act as antagonists (e.g., inverse agonists) ofcomplement C5a receptors, such as human C5a receptors. Within certainaspects, C5a receptor modulators provided herein are1-aryl-4-substituted isoquinolines of Formula I or a pharmaceuticallyacceptable form thereof.

Within certain aspects, compounds provided herein are1-aryl-4-substituted isoquinoline or 1-aryl-3,4-disubstitutedisoquinoline analogues of Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   R₁ is selected from the group consisting of hydrogen, halogen,    cyano, amino, optionally substituted alkyl, optionally substituted    alkenyl, optionally substituted alkynyl, optionally substituted    cycloalkyl, optionally substituted cycloalkenyl, optionally    substituted haloalkyl, optionally substituted haloalkoxy, optionally    substituted alkoxy, optionally substituted cycloalkoxy, optionally    substituted (cycloalkyl)alkoxy, and optionally substituted    heterocycloalkyl;-   R₂ is selected from the group consisting of —NR₄R₅,    —(CR_(A)R_(B))OR₄, —CR_(A)R_(B)NR₄R₅, —C(R_(A′))═CR_(A)R_(B), and    —CR_(A)R_(B)Q;-   R₃ represents between 0 and 4 substituents, each of which is    independently selected from halogen, hydroxy, amino, cyano,    optionally substituted alkyl, optionally substituted haloalkyl,    optionally substituted alkenyl, optionally substituted alkynyl,    optionally substituted cycloalkyl, optionally substituted alkoxy,    optionally substituted haloalkoxy, optionally substituted    hydroxyalkyl, optionally substituted alkoxyalkyl, optionally    substituted mono- and di-alkylamino, optionally substituted    aminoalkyl, -E-(CRcR_(D))_(m)—Z, -E-(CR_(C)R_(D))_(m)—XR_(A);-   R₄ is:    -   (i) C₂-C₈alkyl, C₂-C₈alkenyl, C₂-C₈alkynyl,        (C₃-C₇cycloalkyl)C₀-C₄alkyl, mono- or        di-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl, arylC₀-C₄alkyl, or        (heterocycle)C₀₋₄alkyl, each of which is substituted with from 0        to 4 substituents independently chosen from R_(x),        C₂-C₄alkanoyl, mono- and di-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono-        and di-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl and XR_(y); or    -   (ii) joined to R₅ to form, with the nitrogen to which R₄ and R₅        are bound, a heterocycle having from 1 to 3 rings, 5 to 7 ring        members in each ring, and is substituted with from 0 to 4        substituents independently chosen from R_(X), oxo and W—Z;-   R₅ is:    -   (i) hydrogen;    -   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,        (C₃-C₇carbocycle)C₀-C₄alkyl, each of which is substituted with        from 0 to 3 substituents independently chosen from halogen,        hydroxy, amino, cyano, C₁-C₄alkoxy, methylamino, dimethylamino,        trifluoromethyl and trifluoromethoxy; or    -   (iii) joined to R₄ to form an optionally substituted        heterocycle;-   Ar is mono-, di-, or tri-substituted phenyl, optionally substituted    naphthyl, or optionally substituted heteroaryl, wherein Ar is    optionally substituted heteroaryl when R₂ is —NR₄R₅;-   R_(A), R_(A′), and R_(B), which may be the same or different, are    independently selected at each occurrence from: hydrogen, hydroxy,    and straight or branched alkyl groups, cycloalkyl groups,    (cycloalkyl)alkyl groups and are optionally further substituted with    one or more substituent(s) independently selected from oxo, hydroxy,    halogen, cyano, amino, C₁₋₆alkoxy, —NH(C₁₋₆alkyl),    —N(C₁₋₆alkyl)(C₁₋₆alkyl), —NHC(═O)(C₁₋₆alkyl),    —N(C₁₋₆alkyl)C(═O)(C₁₋₆alkyl), —NHS(O)_(n)(C₁₋₆alkyl),    —S(O)_(n)(C₁₋₆ alkyl), —S(O)_(n)NH(C₁₋₆alkyl); —S(O)_(n)N(C₁₋₆    alkyl)(C₁₋₆alkyl), and Z;-   E is a single bond, oxygen, or NR_(A);-   X is independently selected at each occurrence from the group    consisting of —CH₂—, —CHR_(B)—, —O—, —C(═O)—, —C(═O)O—, —S(O)_(n)—,    —NH—, —NR_(B)—, —C(═O)NH—, —C(═O)NR_(B)—, —S(O)_(n)NH—,    —S(O)_(n)NR_(B)—, —NHC(═O)—, —NR_(B)C(═O)—, —NHS(O)_(n)—, and    —NR_(B)S(O)_(n)—;-   Y and Z are independently selected at each occurrence from 3- to    7-membered carbocyclic or heterocyclic groups which are saturated,    unsaturated, or aromatic, which are optionally substituted with one    or more substituents independently selected from halogen, oxo,    hydroxy, amino, cyano, C₁₋₄alkyl, —O(C₁₋₄alkyl), —NH(C₁₋₄alkyD,    —N(C₁₋₄alkyl)(C₁₋₄alkyl), and —S(O)_(n)(alkyl),-   Q is an optionally substituted carbocyclic or optionally substituted    heterocyclic group which are saturated, unsaturated or aromatic and    comprises between 3 and 18 ring atoms arranged in 1, 2, or 3 rings    which are fused, Spiro or coupled by a bond;-   m is an integer independently selected at each occurrence from    integers in range of 0-8; and-   n is an integer independently selected at each occurrence from 0, 1,    and 2.

Within certain other aspects, compounds provided herein are1-aryl-4-substituted isoquinoline or 1-aryl-3,4-disubstitutedisoquinoline analogues of Formula II:

or a pharmaceutically acceptable form thereof, wherein:

-   Ar is substituted phenyl, optionally substituted naphthyl, or    optionally substituted heteroaryl;-   A is OR₄, NR₄R₅, or CR₄(XR_(y))₂;-   R₁ is chosen from:-   (i) hydrogen, halogen, amino, and cyano; and-   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy,    C₁-C₄haloalkyl, C₁-C₄haloalkoxy, mono- and di-(C₁-C₆alkyl)amino,    (C₃-C₇cycloalkyl)C₀-C₄alkyl, (3- to 7-membered    heterocycloalkyl)C₀-C₄alkyl and —S(O_(n))C₁-C₄alkyl, each of which    is substituted with from 0 to 4 substituents independently chosen    from R_(x);-   R₃ represents between 0 and 4 substituents, each of which is    independently selected from hydrogen, halogen, hydroxy, amino,    cyano, optionally substituted alkyl, optionally substituted alkenyl,    optionally substituted alkynyl, optionally substituted cycloalkyl,    optionally substituted alkoxy, optionally substituted alkoxyalkyl,    optionally substituted hydroxyalkyl, optionally substituted mono-    and di-alkylamino, optionally substituted aminoalkyl, optionally    substituted cycloalkyloxy, optionally substituted aryl, optionally    substituted aralkyl, optionally substituted aryloxy, optionally    substituted aralkyloxy, optionally substituted heterocycle,    optionally substituted heterocycle-oxy, -E-(CR_(C)R_(D))_(m)—Z,    -E-(CR_(C)R_(D))_(m)—XR_(A);-   R₄ is:    -   (i) C₂-C₈alkyl, C₂-C₈alkenyl, C₂-C₈alkynyl,        (C₃-C₇cycloalkyl)C₀-C₄alkyl, mono- or        di-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl, arylC₀-C₄alkyl, or        (heterocycle)C₀₋₄alkyl, each of which is substituted with from 0        to 4 substituents independently chosen from R_(x),        C₂-C₄alkanoyl, mono- and di-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono-        and di-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl and XR_(y); or    -   (ii) joined to R₅ to form, with the nitrogen to which R₄ and R₅        are bound, a heterocycle having from 1 to 3 rings, 5 to 7 ring        members in each ring, and is substituted with from 0 to 4        substituents independently chosen from R_(x), oxo and W—Z;-   R₅ is:    -   (i) hydrogen;    -   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,        (C₃-C₇-carbocycle)C₀-C₄alkyl, each of which is substituted with        from 0 to 3 substituents independently chosen from halogen,        hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy, methylamino,        dimethylamino, trifluoromethyl and trifluoromethoxy; or    -   (iii) joined to R₄ to form an optionally substituted        heterocycle;-   R₈ and R₉ are independently selected from:    -   (i) hydrogen, halogen, hydroxy, C₁-C₆alkyl, C₂-C₆alkenyl,        C₂-C₆alkynyl, C₁-C₆alkoxy, C₁-C₆alkylamino or C₃-C₇cycloalkyl        C₀-C₄alkyl;-   E is a single bond, oxygen, or NR_(A);-   X is a single bond, —CR_(A)R_(B)—, —O—, —C(═O)—, —C(═O)O—,    —S(O)_(n)— or —NR_(B)—; and-   R_(y) is:    -   (i) hydrogen; or    -   (ii) C₁-C₁₀alkenyl, C₂-C₁₀alkynyl, C₃-C₁₀-carbocycleC₀-C₄alkyl        or (3- to 10-membered heterocycle)C₀-C₄alkyl, each of which is        substituted with from 0 to 6 substituents independently selected        from R_(x), oxo, —NH(C₁-C₆alkanoyl),        —N(C₁-C₆alkyl)C₁-C₆alkanoyl, —NHS(O_(n))C₁-C₆alkyl,        —N(S(O_(n))C₁-C₆alkyl)₂, —S(O_(n))NHC₁-C₆alkyl and        —S(O_(n))N(C₁-C₆alkyl)₂;-   W is a single bond, —CR_(A)R_(B)—, —NR_(B)— or —O—;-   Z is independently selected at each occurrence from 3- to 7-membered    carbocycles and heterocycles, each of which is substituted with from    0 to 4 substituents independently selected from halogen, oxo, —COOH,    hydroxy, amino, cyano, C₁-C₆alkyl, C₁-C₆alkoxy, C₆haloalkyl,    C₁-C₆haloalkoxy, mono- and di-(C₁-C₆alkyl)amino and    —S(O_(n))C₁-C₆alkyl; and-   R_(A) and R_(B) are independently selected at each occurrence from:    -   (i) hydrogen; and    -   (ii) C₁-C₁₀alkyl, C₂-C₁₀alkenyl, C₂-C₁₀alkynyl, saturated or        partially saturated (C₃-C₁₀carbocycle)C₀-C₄alkyl and saturated        or partially saturated (3- to 10-membered        heterocycle)C₀-C₄alkyl, each of which is substituted with from 0        to 6 substituents independently selected from oxo, hydroxy,        halogen, cyano, amino, C₁-C₆alkoxy, mono- and        di-(C₁-C₄alkyl)amino, —COOH, —C(═O)NH₂, —NHC(═O)(C₁-C₆alkyl),        —N(C₁-C₆alkyl)C(═O)(C₁-C₆alkyl), —NHS(O_(n))C₁-C₆alkyl, SO₃H,        —S(O_(n))C₁-C₆alkyl, —S(O_(n))NHC₁-C₆alkyl,        —S(O_(n))N(C₁-C₆alkyl)C₁-C₆alkyl and Z;-   R_(C) and R_(D) are independently selected from R_(A), hydroxy,    C₁₋₆alkoxy, and oxo;-   R_(x) is independently chosen at each occurrence from halogen,    hydroxy, amino, cyano, nitro, —COOH, —C(═O)NH₂, C₁-C₆alkoxycarbonyl,    —C(═O)NHC₁-C₆alkyl, —C(═O)N(C₁-C₆alkyl)₂, C₁-C₆alkyl, C₂-C₆alkenyl,    C₂-C₆alkynyl, mono- and di-(C₁-C₆alkyl)amino, C₁-C₆alkoxy,    C₁-C₂hydroxyalkyl, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,    (C₃-C₇cycloalkyl)C₀-C₄alkyl, and —S(O_(n))C₁-C₆alkyl;-   m is an integer independently selected at each occurrence from 0-8;    and-   n is an integer independently selected at each occurrence from 0, 1    and 2.

Within certain other aspects, compounds provided herein are1-aryl-4-substituted isoquinoline or 1-aryl-3,4-disubstitutedisoquinoline analogues of Formula IX:

or a pharmaceutically acceptable salt thereof, wherein:

-   R₁ is selected from the group consisting of hydrogen, halogen,    cyano, amino, optionally substituted alkyl, optionally substituted    alkenyl, optionally substituted alkynyl, optionally substituted    cycloalkyl, optionally substituted cycloalkenyl, optionally    substituted haloalkyl, optionally substituted haloalkoxy, optionally    substituted alkoxy, optionally substituted cycloalkoxy, optionally    substituted (cycloalkyl)alkoxy, and optionally substituted    heterocycloalkyl;-   R₃ represents between 0 and 4 substituents, each of which is    independently selected from optionally substituted alkyl, optionally    substituted alkenyl, optionally substituted alkynyl, optionally    substituted alkoxy, optionally substituted hydroxyalkyl, optionally    substituted aminoalkyl, optionally substituted mono- and    di-alkylamino, —O—(CR_(A)R_(B))_(m)—XR_(A), —O—(CR_(A)R_(B))_(m)—Y,    —N(R_(B))—(CR_(A)R_(B))_(m)—XR_(A), —N(R_(B))—(CR_(A)R_(B))_(m)—Y;-   R₄ is:    -   (i) C₂-C₈alkyl, C₂-C₈alkenyl, C₂-C₈alkynyl,        (C₃-C₇cycloalkyl)C₀-C₄alkyl, mono- or        di-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl, arylC₀-C₄alkyl, or        (heterocycle)C₀₋₄alkyl, each of which is substituted with from 0        to 4 substituents independently chosen from R_(x),        C₂-C₄alkanoyl, mono- and di-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono-        and di-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl and XR_(y); or    -   (ii) joined to R₅ to form, with the nitrogen to which R₄ and R₅        are bound, a heterocycle having from 1 to 3 rings, 5 to 7 ring        members in each ring, and is substituted with from 0 to 4        substituents independently chosen from R_(x), oxo and W—Z;-   R₅ is:    -   (i) hydrogen;    -   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,        (C₃-C₇carbocycle)C₀-C₄alkyl, each of which is substituted with        from 0 to 3 substituents independently chosen from halogen,        hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy, methylamino,        dimethylamino, trifluoromethyl and trifluoromethoxy; or    -   (iii) joined to R₄ to form an optionally substituted        heterocycle;-   Ar is optionally substituted naphthyl or optionally substituted    heteroaryl;-   R_(A) and R_(B), which may be the same or different, are    independently selected at each occurrence from: hydrogen, hydroxy,    and straight or branched alkyl groups, cycloalkyl groups,    (cycloalkyl)alkyl groups and are optionally further substituted with    one or more substituent(s) independently selected from oxo, hydroxy,    halogen, cyano, amino, C₁₋₆alkoxy, —NH(C₁₋₆alkyl),    —N(C₁₋₆alkyl)(C₁₋₆alkyl), —NHC(═O)(C₁₋₆alkyl),    —N(C₁₋₆alkyl)C(═O)(C₁₋₆alkyl), —NHS(O)_(n)(C₁₋₆alkyl),    —S(O)_(n)(C₁₋₆ alkyl), —S(O)_(n)NH(C₁₋₆alkyl), —S(O)_(n)N(C₁₋₆    alkyl)(C₁₋₆alkyl), and Z;-   X is independently selected at each occurrence from the group    consisting of —CH₂—, —CHR_(B)—, —O—, —C(═O)—, —C(═O)O—, —S(O)_(n)—,    —NH—, —NR_(B)—, —C(═O)NH—, —S(O)_(n)NH—, —S(O)_(n)NR_(B)—,    —NHC(═O)—, —NR_(B)C(═O)—, —NHS(O)_(n)—, and —NR_(B)S(O)_(n)—;-   Y and Z are independently selected at each occurrence from 3- to    7-membered carbocyclic or heterocyclic groups which are saturated,    unsaturated, or aromatic, which are optionally substituted with one    or more substituents independently selected from halogen, oxo,    hydroxy, amino, cyano, C₁₋₄alkyl, —O(C₁₋₄alkyl), —NH(C₁₋₄alkyl),    —N(C₁₋₄alkyl)(C₁₋₄alkyl), and —S(O)_(n)(alkyl),-   m is an integer independently selected at each occurrence from    integers in range of 0-8; and-   n is an integer independently selected at each occurrence from 0, 1,    and 2.

In certain embodiments, C5a receptor modulators provided herein exhibithigh affinity for C5a receptor (i.e., an affinity constant for bindingto C5a receptor of less than 1 micromolar) or very high affinity for C5areceptor (i.e., an affinity constant for binding to the C5a receptor ofless than 100 nanomolar). In certain embodiments, such modulatorsexhibit an affinity for human C5a receptor that is higher than for rator mouse C5a receptor, preferably at least five times higher, morepreferably ten times higher. Affinity of a compound for C5a receptor maybe determined, for example, via a radioligand binding assay, such as theassay provided in Example 60.

Within certain aspects, modulators as described herein are C5a receptorantagonists, such as inverse agonists. Certain such compounds exhibit anEC₅₀ of 1 micromolar or less, 500 nM or less, 100 nM or less, or 25 nMor less, in a standard in vitro C5a receptor-mediated chemotaxis assay(such as the assay provided in Example 55) or a calcium mobilizationassay (as described in Example 62).

Within further aspects, C5a receptor antagonists are essentially free ofC5a receptor agonist activity (i.e., exhibit less than 5% agonistactivity in a GTP binding assay as described in Example 61).

The present invention further provides, within other aspects,pharmaceutical compositions comprising at least one C5a receptormodulator as described herein, in combination with a physiologicallyacceptable carrier or excipient. Processes for preparing suchpharmaceutical compositions are also provided. Such compositions areparticularly useful in the treatment of C5a-mediated inflammation, suchas inflammation associated with various inflammatory and immune systemdisorders.

Within further aspects, methods are provided for inhibitingsignal-transducing activity of a cellular C5a receptor, comprisingcontacting a cell expressing a C5a receptor with at least one C5areceptor modulator as described herein, and thereby reducing signaltransduction by the C5a receptor.

Methods are further provided for inhibiting binding of C5a to C5areceptor in vitro, comprising contacting C5a receptor with at least oneC5a receptor modulator as described herein, under conditions and in anamount sufficient to detectably inhibit C5a binding to C5a receptor.

The present invention further provides methods for inhibiting binding ofC5a to C5a receptor in a human patient, comprising contacting cellsexpressing C5a receptor with at least one C5a receptor modulator asdescribed herein.

Within further aspects, the present invention provides methods fortreating a patient in need of anti-inflammatory treatment orimmunomodulatory treatment. Such methods generally compriseadministering to the patient a C5a receptor modulatory amount of a C5areceptor modulator as described herein. Treatment of humans,domesticated companion animals (pets) or livestock animals sufferingsuch conditions is contemplated by the present invention. In certainsuch aspects, methods are provided for treating a patient suffering fromcystic fibrosis, rheumatoid arthritis, psoriasis, cardiovasculardisease, reperfusion injury, or bronchial asthma comprisingadministering to the patient a C5a receptor modulatory amount of a C5areceptor modulator as described herein. In further such aspects, methodsare provided for treating a patient suffering from stroke, myocardialinfarction, atherosclerosis, ischemic heart disease, orischemia-reperfusion injury comprising administering to the patient aC5a receptor modulatory amount of a C5a receptor modulator as describedherein.

The present invention further provides methods for inhibiting C5areceptor-mediated cellular chemotaxis (preferably leukocyte (e.g.,neutrophil) chemotaxis), comprising contacting mammalian white bloodcells with a C5a receptor modulatory amount of a C5a receptor modulatoras described herein. In certain embodiments, the white blood cells areprimate white blood cells, such as human white blood cells.

Within further aspects, the present invention provides methods for usinga C5a receptor modulator as described herein as a probe for thelocalization of receptors, particularly C5a receptors. Such localizationmay be achieved, for example, in tissue sections (e.g., viaautoradiography) or in vivo (e.g., via positron emission tomography,PET, or single positron emission computed tomography, SPECT, scanningand imaging). Within certain such aspects, the present inventionprovides methods for localizing C5a receptors in a tissue sample,comprising: (a) contacting the tissue sample containing C5a receptorswith a detectably labeled compound as described herein under conditionsthat permit binding of the compound to C5a receptors; and (b) detectingthe bound compound. Such methods may, optionally, further comprise astep of washing the contacted tissue sample, prior to detection.Suitable detectable labels include, for example, radiolabels such as¹²⁵I, tritium, ¹⁴C, ³²P and ⁹⁹Tc.

The present invention also provides packaged pharmaceuticalpreparations, comprising: (a) a pharmaceutical composition as describedherein in a container; and (b) instructions for using the composition totreat a patient suffering from one or more conditions responsive to C5areceptor modulation, such as rheumatoid arthritis, psoriasis,cardiovascular disease, reperfusion injury, bronchial asthma, stroke,myocardial infarction, atherosclerosis, ischemic heart disease, orischemia-reperfusion injury.

In yet another aspect, the present invention provides methods forpreparing the compounds disclosed herein, including the intermediates.

These and other aspects of the present invention will become apparentupon reference to the following detailed description.

DETAILED DESCRIPTION OF THE INVENTION

As noted above, the present invention provides 1-aryl-4-substitutedisoquinolines that modulate C5a receptor activation and/or C5areceptor-mediated signal transduction. Such compounds may be used invitro or in vivo to modulate (preferably inhibit) C5a receptor activityin a variety of contexts.

Chemical Description and Terminology

Compounds provided herein are generally described using standardnomenclature. For compounds having asymmetric centers, it should beunderstood that (unless otherwise specified) all of the optical isomersand mixtures thereof are encompassed. Compounds with two or moreasymmetric elements can also be present as mixtures of diastereomers. Inaddition, compounds with carbon-carbon double bonds may occur in Z- andE-forms, with all isomeric forms of the compounds being included in thepresent invention unless otherwise specified. Where a compound exists invarious tautomeric forms, a recited compound is not limited to any onespecific tautomer, but rather is intended to encompass all tautomericforms. Recited compounds are further intended to encompass compounds inwhich one or more atoms are replaced with an isotope (i.e., an atomhaving the same atomic number but a different mass number). By way ofgeneral example, and without limitation, isotopes of hydrogen includetritium and deuterium and isotopes of carbon include ¹¹C, ¹³C, and ¹⁴C.

Certain compounds are described herein using a general formula thatincludes variables (e.g., R₁—R₅, R₈—R₁₃, Ar). Unless otherwisespecified, each variable within such a formula is defined independentlyof any other variable, and any variable that occurs more than one timein a formula is defined independently at each occurrence. Thus, forexample, if a group is shown to be substituted with 0-2 R*, the groupmay be unsubstituted or substituted with up to two R* groups and R* ateach occurrence is selected independently from the definition of R*.Also, combinations of substituents and/or variables are permissible onlyif such combinations result in stable compounds.

The term “1-aryl-4-substituted isoquinolines,” as used herein, refers tocompounds of Formula I, as well as pharmaceutically acceptable formsthereof. Such compounds may, but need not, further satisfy one or moreadditional Formulas provided herein.

“Pharmaceutically acceptable forms” of the compounds recited hereininclude pharmaceutically acceptable salts, esters, hydrates, clathratesand prodrugs of such compounds. As used herein, a pharmaceuticallyacceptable salt is an acid or base salt that is generally considered inthe art to be suitable for use in contact with the tissues of humanbeings or animals without excessive toxicity, irritation, allergicresponse, or other problem or complication. Such salts include mineraland organic acid salts of basic residues such as amines, as well asalkali or organic salts of acidic residues such as carboxylic acids.Specific pharmaceutical salts include, but are not limited to, salts ofacids such as hydrochloric, phosphoric, hydrobromic, malic, glycolic,fumaric, sulfuric, sulfamic, sulfanilic, formic, toluenesulfonic,methanesulfonic, benzene sulfonic, ethane disulfonic,2-hydroxyethylsulfonic, nitric, benzoic, 2-acetoxybenzoic, citric,tartaric, lactic, stearic, salicylic, glutamic, ascorbic, pamoic,succinic, fumaric, maleic, propionic, hydroxymaleic, hydroiodic,phenylacetic, alkanoic such as acetic, HOOC—(CH₂)_(n)—COOH where n is0-4, and the like. Similarly, pharmaceutically acceptable cationsinclude, but are not limited to sodium, potassium, calcium, aluminum,lithium and ammonium. Those of ordinary skill in the art will recognizefurther pharmaceutically acceptable salts for the compounds providedherein, including those listed by Remington's Pharmaceutical Sciences,17th ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985).

A “prodrug” is a compound that may not fully satisfy the structuralrequirements of Formula I (or another Formula as provided herein) but ismodified in vivo, following administration to a patient, to produce sucha compound. For example, a prodrug may be an acylated derivative of acompound as provided herein. Prodrugs include compounds wherein hydroxy,amine or sulfhydryl groups are bonded to any group that, whenadministered to a mammalian subject, cleaves to form a free hydroxyl,amino, or sulfhydryl group, respectively. Examples of prodrugs include,but are not limited to, acetate, formate and benzoate derivatives ofalcohol and amine functional groups within the compounds providedherein.

A “C5a receptor modulatory amount” is an amount that, uponadministration, results in a concentration of C5a receptor modulator ata C5a receptor that is sufficient to inhibit chemotaxis of white bloodcells in an in vitro assay and/or alter C5a receptor activity oractivation as measured by an in vitro calcium mobilization assay. In achemotaxis assay (see Example 55), the level of C5a-induced chemotaxisobserved in a control assay (i.e., one to which a compound as providedherein has not been added) is significantly higher (measured as p<0.05using a conventional parametric statistical analysis method such as astudent's T-test) than the level observed in an assay to which acompound or form thereof as described herein has been added. Within suchan assay, the C5a is generally from the same species as the cells usedin the assay. In a calcium mobilization assay (see Example 62), aconcentration of compound that alters C5a receptor activity oractivation may inhibit C5a-induced calcium mobilization or may itselfincrease or decrease C5a receptor-mediated calcium mobilization in theabsence of C5a.

A “therapeutically effective amount” is an amount of a compound or formthereof as provided herein that, upon administration, results in adiscernible benefit in a patient. Such benefit may be confirmed usingstandard clinical procedures.

A “substituent,” as used herein, refers to a molecular moiety that iscovalently bonded to an atom within a molecule of interest. For example,a “ring substituent” may be a moiety such as a halogen, alkyl group,haloalkyl group or other substituent described herein that is covalentlybonded to an atom (preferably a carbon or nitrogen atom) that is a ringmember. The term “substituted,” as used herein, means that any one ormore hydrogens on the designated atom is replaced with a selection fromthe indicated substituents, provided that the designated atom's normalvalence is not exceeded, and that the substitution results in a stablecompound (i.e., a compound that can be isolated, characterized andtested for biological activity). When a substituent is oxo (i.e., =0),then 2 hydrogens on the atom are replaced. When aromatic moieties aresubstituted by an oxo group, the aromatic ring is replaced by thecorresponding partially unsaturated ring. For example a pyridyl groupsubstituted by oxo is a pyridone.

The phrase “optionally substituted” indicates that a group may either beunsubstituted or substituted at one or more of any of the availablepositions, typically 1, 2, 3, 4, or 5 positions, by one or more suitablesubstituents such as those disclosed herein. Optional substitution mayalso be indicated by the phrase “substituted with from 0 to Xsubstituents,” in which X is the maximum number of substituents.

Suitable substituents include, for example, halogen, cyano, amino,hydroxy, nitro, azido, carboxamido, —COOH, SO₂NH₂, alkyl (e.g.,C₁-C₈alkyl), alkenyl (e.g., C₂-C₈alkenyl), alkynyl (e.g., C₂-C₈alkynyl),alkoxy (e.g., C₁-C₈alkoxy), alkyl ether (e.g., C₂-C₈alkyl ether),alkylthio (e.g., C₁-C₈alkylthio), haloalkyl (e.g., C₁-C₈haloalkyl),hydroxyalkyl (e.g., C₁-C₈hydroxyalkyl), aminoalkyl (e.g.,C₁-C₈aminoalkyl), haloalkoxy (e.g., C₁-C₈haloalkoxy), alkanoyl (e.g.,C₁-C₈alkanoyl), alkanone (e.g., C₁-C₈alkanone), alkanoyloxy (e.g.,C₁-C₈alkanoyloxy), alkoxycarbonyl (e.g., C₁-C₈alkoxycarbonyl), mono- anddi-(C₁-C₈alkyl)amino, mono- and di-(C₁-C₈alkyl)aminoC₁-C₈alkyl, mono-and di-(C₁-C₈alkyl)carboxamido, mono- and di-(C₁-C₈alkyl)sulfonamido,alkylsulfinyl (e.g., C₁-C₈alkylsulfinyl), alkylsulfonyl (e.g.,C₁-C₈alkylsulfonyl), aryl (e.g., phenyl), arylalkyl (e.g.,(C₆-C₁₈aryl)C₁-C₈alkyl, such as benzyl and phenethyl), aryloxy (e.g.,C₆-C₁₈aryloxy such as phenoxy), arylalkoxy (e.g.,(C₆-C₁₈aryl)C₁-C₈alkoxy) and/or 3- to 8-membered heterocyclic groupssuch as coumarinyl, quinolinyl, pyridyl, pyrazinyl, pyrimidyl, furyl,pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, indolyl,benzofuranyl, benzothiazolyl, tetrahydropyranyl, tetrahydropyranyl,piperidinyl, morpholino or pyrrolidinyl. Certain groups within theformulas provided herein are optionally substituted with from 1 to 3, 1to 4 or 1 to 5 independently selected substituents.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example, —CONH₂ isattached through the carbon atom.

As used herein, “alkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups, and wherespecified, having the specified number of carbon atoms. Thus, the termC₁-C₆alkyl, as used herein, indicates an alkyl group having from 1 to 6carbon atoms. “C₀-C₄alkyl” refers to a bond or a C₁-C₄alkyl group. Alkylgroups include groups having from 1 to 8 carbon atoms (C₁-C₈alkyl), from1 to 6 carbon atoms (C₁-C₆alkyl) and from 1 to 4 carbon atoms(C₁-C₄alkyl), such as methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl,2-hexyl, 3-hexyl, and 3-methylpentyl. In certain embodiments, preferredalkyl groups are methyl, ethyl, propyl, butyl, and 3-pentyl.“Aminoalkyl” is an alkyl group as defined herein substituted with one ormore —NH₂ substituents. “Hydroxyalkyl” is a hydroxy group as definedherein substituted with one or more —OH substituents.

“Alkenyl” refers to a straight or branched hydrocarbon chain comprisingone or more unsaturated carbon-carbon bonds, such as ethenyl andpropenyl. Alkenyl groups include C₂-C₈alkenyl, C₂-C₆alkenyl andC₂-C₄alkenyl groups (which have from 2 to 8, 2 to 6 or 2 to 4 carbonatoms, respectively), such as ethenyl, allyl or isopropenyl.

“Alkynyl” refers to straight or branched hydrocarbon chains comprisingone or more triple carbon-carbon bonds. Alkynyl groups includeC₂-C₈alkynyl, C₂-C₆alkynyl and C₂-C₄alkynyl groups, which have from 2 to8, 2 to 6 or 2 to 4 carbon atoms, respectively. Alkynyl groups includefor example groups such as ethynyl and propynyl.

By “alkoxy,” as used herein, is meant an alkyl, alkenyl or alkynyl groupas described above attached via an oxygen bridge. Alkoxy groups includeC₁-C₆alkoxy and C₁-C₄alkoxy groups, which have from 1 to 6 or 1 to 4carbon atoms, respectively. Methoxy, ethoxy, propoxy, isopropoxy,n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy,isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxyare specific alkoxy groups. Similarly “alkylthio” refers to an alkyl,alkenyl or alkynyl group as described above attached via a sulfurbridge.

The term “alkanoyl” refers to an alkyl group as defined above attachedthrough a carbonyl bridge. Alkanoyl groups include C₂-C_(g)alkanoyl,C₂-C₆alkanoyl and C₂-C₄alkanoyl groups, which have from 2 to 8, 2 to 6or 2 to 4 carbon atoms, respectively. “C₁alkanoyl” refers to —(C═O)—H,which (along with C₂-C₈alkanoyl) is encompassed by the term“C₁-C₈alkanoyl.” Ethanoyl is C₂alkanoyl.

An “alkanone” is an alkyl group as defined above with the indicatednumber of carbon atoms substituted at least one position with an oxogroup. “C₃-C₈alkanone,” “C₃-C₆alkanone” and “C₃-C₄alkanone” refer to analkanone having from 3 to 8, 6 or 4 carbon atoms, respectively. By wayof example, a C₃ alkanone group has the structure —CH₂—(C═O)—CH₃.

Similarly, “alkyl ether” refers to a linear or branched ethersubstituent linked via a carbon-carbon bond. Alkyl ether groups includeC₂-C₈alkyl ether, C₂-C₆alkyl ether and C₂-C₄alkyl ether groups, whichhave 2 to 8, 6 or 4 carbon atoms, respectively. By way of example, a C₂alkyl ether group has the structure —CH₂—O—CH₃.

The term “alkoxycarbonyl” refers to an alkoxy group linked via acarbonyl (i.e., a group having the general structure —C(═O)—O-alkyl).Alkoxycarbonyl groups include C₂-C₈, C₂-C₆ and C₂-C₄alkoxycarbonylgroups, which have from 2 to 8, 6 or 4 carbon atoms, respectively.

“Alkanoyloxy,” as used herein, refers to an alkanoyl group linked via anoxygen bridge (e.g., a group having the general structure—O—C(═O)-alkyl). Alkanoyloxy groups include C₂-C₈, C₂-C₆ andC₂-C₄alkanoyloxy groups, which have from 2 to 8, 6 or 4 carbon atoms,respectively.

“Alkylamino” refers to a secondary or tertiary amine having the generalstructure —NH-alkyl or —N(alkyl)(alkyl), wherein each alkyl may be thesame or different. Such groups include, for example, mono- anddi-(C₁-C₈alkyl)amino groups, in which each alkyl may be the same ordifferent and may contain from 1 to 8 carbon atoms, as well as mono- anddi-(C₁-C₆alkyl)amino groups and mono- and di-(C₁-C₄alkyl)amino groups.“Mono- or di-(C₁-C₄alkylamino)C₀-C₄alkyl” refers to a mono- anddi-(C₁-C₄alkyl)amino group that is linked via a direct bond or a C₁-C₄alkyl group (i.e., a group having the general structure—C₀-C₄alkyl-NH-alkyl or —C₀-C₄alkyl-N(alkyl)(alkyl), in which each alkylmay be the same or different. Similarly, “alkylaminoalkoxy” refers to analkylamino group linked via an alkoxy group.

The term “aminocarbonyl” or “carboxamido” refers to an amide group(i.e., —(C═O)NH₂). “Mono- or di-(C₁-C₆alkyl)aminocarbonyl” refers to anamide group in which one or both of the hydrogen atoms is replaced withan independently chosen C₁-C₆alkyl. Such groups may also be indicated by“—C(═O)NHalkyl” or “—C(═O)N(alkyl)alkyl.”

The term “halogen” refers to fluorine, chlorine, bromine and iodine. A“haloalkyl” is a branched or straight-chain alkyl group, substitutedwith 1 or more halogen atoms (e.g., “haloC₁-C₈alkyl” groups have from 1to 8 carbon atoms; “haloC₁-C₆alkyl” groups have from 1 to 6 carbonatoms). Examples of haloalkyl groups include, but are not limited to,mono-, di- or tri-fluoromethyl; mono-, di- or tri-chloromethyl; mono-,di-, tri-, tetra- or penta-fluoroethyl; and mono-, di-, tri-, tetra- orpenta-chloroethyl. Typical haloalkyl groups are trifluoromethyl anddifluoromethyl. Within certain compounds provided herein, not more than5 or 3 haloalkyl groups are present. The term “haloalkoxy” refers to ahaloalkyl group as defined above attached via an oxygen bridge.“HaloC₁-C₈alkoxy” groups have 1 to 8 carbon atoms.

A “carbocycle” is any saturated, partially saturated, or aromatic grouphaving 1 or 2 fused, pendant or Spiro rings, with 3 to 8 atoms in eachring, and with all ring members being carbon. The term “carbocycle”encompasses aromatic groups such as phenyl and naphthyl, as well asgroups that comprise both aromatic and nonaromatic rings (e.g.,tetrahydronaphthyl), and groups with saturated and partially saturatedrings (such as cyclohexyl and cyclohexenyl). When substitutions areindicated, carbocycles may be substituted on any ring atom where suchsubstitution results in a stable compound. The term “C₃-C₁₀carbocycle”refers to such groups having from 3 to 10 ring members. A“C₃-C₁₀carbocycleC₀-C₄alkyl” group is a C₃-C₁₀carbocycle that is linkedvia a direct bond or a C₁-C₄alkyl group.

Certain carbocycles are “cycloalkyl” (i.e., a saturated or partiallysaturated carbocycle). Such groups typically contain from 3 to about 8ring carbon atoms; in certain embodiments, such groups have from 3 to 7ring carbon atoms. Examples of cycloalkyl groups include cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl, as well as such groups modifiedby the presence of one or more double or triple bonds (e.g.,cyclohexenyl) and bridged or caged saturated ring groups such asnorbornane or adamantane. If substituted, any ring carbon atom may bebonded to any indicated substituent.

In the term “(cycloalkyl)alkyl”, “cycloalkyl” and “alkyl” are as definedabove, and the point of attachment is on the alkyl group. This termencompasses, but is not limited to, cyclopropylmethyl, cyclohexylmethyland cyclohexylethyl. “(C₃-C₇cycloalkyl)C₀-C₄alkyl” refers to 3- to7-membered cycloalkyl rings that are linked via a direct bond or aC₁-C₄alkyl.

Other carbocycles are “aryl” (i.e., carbocycles that comprise at leastone aromatic ring). In addition to the aromatic ring(s), additionalnon-aromatic ring(s) may be present in an aryl group. Representativearyl groups include phenyl, naphthyl (e.g., 1-naphthyl and 2-naphthyl),biphenyl, tetrahydronaphthyl and indanyl.

The term “arylalkyl” refers to an aryl group that is linked via an alkylgroup. Certain arylalkyl groups are arylC₀-C₂alkyl, in which an arylgroup is linked via a direct bond or a methylene or ethylene moiety.Such groups include, for example, groups in which phenyl or naphthyl islinked via a bond or C₁-C₂alkyl, such as benzyl, 1-phenyl-ethyl and2-phenyl-ethyl.

The term “aryloxy” refers to an aryl group linked via a an oxygen (i.e.,a group having the general structure —O-aryl). Phenoxy is arepresentative aryloxy group.

A “heteroatom” is an atom other than carbon, such as oxygen, sulfur ornitrogen.

The term “heterocycle” or “heterocyclic group” is used to indicatesaturated, partially unsaturated, or aromatic groups having 1 or 2fused, pendent or Spiro rings, with 3 to 8 atoms in each ring, and in atleast one ring from 1 to 4 heteroatoms independently selected from N, Oand S, with remaining atoms being carbon. Certain heterocycles are 3- to10-membered monocyclic or bicyclic groups; other are 4- to 6-memberedmonocyclic groups. The heterocyclic ring may be attached at anyheteroatom or carbon atom that results in a stable structure, and may besubstituted on carbon and/or nitrogen atom(s) if the resulting compoundis stable. Any nitrogen and/or sulfur heteroatoms may optionally beoxidized, and any nitrogen may optionally be quaternized.

Variations on the term “(heterocycle)alkyl” refer to a heterocycle thatis linked via a direct bond or alkyl group. Such groups include, forexample, (3- to 10-membered heterocycle)C₀-C₄alkyl groups, in which theheterocycle contains from 3 to 10 ring members and is linked via adirect bond or C₁-C₄alkyl. Unless otherwise specified, the heterocycleportion of such groups may be saturated, partially saturated oraromatic. “(4- to 6-membered heterocycloalkyl)C₀-C₄alkyl” refers to aheterocycloalkyl group of 4 to 6 ring members that is linked via adirect bond or a C₁-C₄alkyl.

Certain heterocycles are “heteroaryl” (i.e., groups that comprise atleast one aromatic ring having from 1 to 4 heteroatoms). When the totalnumber of S and 0 atoms in a heteroaryl group exceeds 1, then theseheteroatoms are not adjacent to one another; preferably the total numberof S and 0 atoms in a heteroaryl is not more than 1, 2 or 3, morepreferably 1 or 2 and most preferably not more than 1. Examples ofheteroaryl groups include pyridyl, furanyl, indolyl, pyrimidinyl,pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, thienyl, thiazolyl,triazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, and5,6,7,8-tetrahydroisoquinoline.

Other heterocycles are referred to herein as “heterocycloalkyl” (i.e.,saturated or partially saturated heterocycles). Heterocycloalkyl groupshave 1 or 2 rings, each with from 3 to about 8 ring atoms, and moretypically from 5 to 7 ring atoms. Examples of heterocycloalkyl groupsinclude morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl.

Additional examples of heterocyclic groups include, but are not limitedto, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl,benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl,carbazolyl, NH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl,decahydroquinolinyl, 2H, 6H-1,5,2-dithiazinyl,dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl,indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl,isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl,isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl,oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl,1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl,phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl,pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl,pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole,pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl,pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl,quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl,tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4thiadiazolyl, thianthrenyl,thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl,thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl,1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl.

“A C5a receptor” is a G-protein coupled receptor that specifically bindsC5a peptide. Certain preferred C5a receptors are human, such as theprotein product of the sequence that produces the human C5a receptor PCRproduct described by Gerard and Gerard (1991) Nature 349:614-17. Thehuman C5a receptor may also be that described by Boulay (1991)Biochemistry 30(12):2993-99 (nucleotide sequence encoding the receptoris available at GENBANK Accession No. M62505). Non-primate C5a receptorsinclude the rat C5a receptor (encoded by the nucleotide sequence havingGENBANK Accession No. X65862, Y09613 or AB003042), canine C5a receptor(encoded by the nucleotide sequence having GENBANK Accession No.X65860), and guinea pig C5a receptor (encoded by the nucleotide sequencehaving GENBANK Accession No. U86103).

A “C5a receptor modulator” (also referred to herein as a “modulator”) isany compound that modulates C5a receptor activation and/or activity(i.e., C5a receptor-mediated signal transduction, as measured using aC5a receptor-mediated chemotaxis, radioligand binding assay, or calciummobilization assay as provided herein). In certain embodiments, such amodulator may be exhibit an affinity constant for binding to a C5areceptor of less than 1 micromolar in a standard C5a receptorradioligand binding assay; and/or an EC₅₀ of less than 1 micromolar in astandard C5a receptor-mediated chemotaxis assay or calcium mobilizationassay. In other embodiments the a C5a receptor modulator may exhibit anaffinity constant or EC₅₀ of less than 500 nM, 200 nM, 100 nM, 50 nM, 25nM, 10 nM or 5 nM in such an assay. A modulator may be a C5a receptoragonist or antagonist, although, for certain purposes described herein,a modulator preferably inhibits C5a activation resulting from binding ofC5a (i.e., the modulator is an antagonist). In addition, oralternatively, a modulator may act as an inverse agonist of C5areceptor. In certain embodiments, modulators provided herein modulateactivation and/or activity of a primate C5a receptor, such as human C5areceptor, which may be a cloned, recombinantly expressed receptor or anaturally expressed receptor. For treating non-human animals of anyparticular species, a compound exhibiting high affinity for the C5areceptor of that particular species is preferred.

Certain C5a receptor modulators exhibit high activity in a standard invitro C5a receptor mediated chemotaxis assay, as specified in Example55, herein. Such compounds exhibit an EC₅₀ of 4 μM or less in such astandard C5a mediated chemotaxis assay, preferably an EC₅₀ of 1 μM orless in such an assay, more preferably an EC₅₀ of 0.1 μM or less in suchan assay, and even more preferably and EC₅₀ of 10 nM or less in such anassay.

An “inverse agonist” of a C5a receptor is a compound that reduces theactivity of the C5a receptor below its basal activity level in theabsence of added C5a. Inverse agonists may also inhibit the activity ofC5a at the C5a receptor, and/or may inhibit binding of C5a to the C5areceptor. The ability of a compound to inhibit the binding of C5a to theC5a receptor may be measured by a binding assay, such as the radioligandbinding assay given in Example 60. The basal activity of the C5areceptor may be determined from a GTP binding assay, such as the assayof Example 61. The reduction of C5a receptor activity may also bedetermined from a GTP binding assay or a calcium mobilization assay suchas the assay of Example 62.

A “neutral antagonist of the C5a receptor is a compound which inhibitsthe activity of C5a at the C5a receptor, but does not significantlychange the basal activity of the C5a receptor. Neutral antagonists ofthe C5a receptor may inhibit the binding of C5a to the C5a receptor.

A “partial agonist” of the C5a receptor elevates the activity of the C5areceptor above the basal activity level of the receptor in the absenceof C5a, but does not elevate the activity of the C5a receptor to thelevel brought about by saturating levels of the natural agonist, C5a.Partial agonist compounds may inhibit the binding of C5a to the C5areceptor. Partial agonists of the C5a receptor usually elevate theactivity of the C5a receptor, producing a level of elevation rangingfrom 5% to 90% of the activity level brought about byreceptor-saturating concentrations of the natural agonist, C5a.

C5A Receptor Modulators

As noted above, the present invention provides C5a receptor modulators.Such modulators may be used to alter C5a receptor activity in a varietyof contexts, including in the treatment of patients suffering fromdiseases or disorders responsive to C5a receptor modulation, such asautoimmune disorders and inflammatory conditions. C5a receptormodulators may also be used within a variety of in vitro assays (e.g.,assays for receptor activity), as probes for detection and localizationof C5a receptor and as standards in assays of ligand binding and C5areceptor-mediated signal transduction.

C5a receptor modulators provided herein are 1-aryl-4-substitutedisoquinolines of Formula I (as well as pharmaceutically acceptable formsthereof) that detectably alter, preferably decrease, C5a receptoractivation and/or signal transduction activity at submicromolarconcentrations.

Certain preferred compounds of Formula II, include those compounds inwhich R₁ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,C₁-C₆alkoxy, C₁-C₆haloalkyl, C₁-C₆haloalkoxy,(C₃-C₇cycloalkyl)-C₀-C₄alkyl. Other preferred compounds of Formula IIinclude those compounds in which R₁ is hydrogen, C₁-C₄alkyl orC₁-C₄alkoxy, or more preferably R₁ is hydrogen, methyl, ethyl, ormethoxy.

Other preferred compounds of Formula II, include those compounds inwhich R₃ represents between 0 and 2 substituents, each of which isindependently selected from C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,C₁₋₆haloalkoxy, hydroxy, COOH, CONH₂, SO₂NH₂, mono- anddi-(C₁₋₆alkyl)amino, (amino)C₀₋₆alkyl. In certain other compounds R₃ ispreferably absent, e.g., the carbocyclic ring of the isoquinoline isunsubstituted. Other preferred compounds of Formula II include thosecompounds in which R₃ is R₃ represents between 0 and 2 substituents,each of which is independently selected from C₁₋₆alkyl, C₁₋₆alkoxy,hydroxy, COOH, CONH₂, and SO₂NH₂.

Yet other preferred compounds of Formula II, include those compounds inwhich R₄ is:

-   -   (i) C₂-C₈alkyl, C₂-C₈alkenyl, C₂-C₈alkynyl,        (C₃-C₇cycloalkyl)C₀-C₄alkyl, mono- or        di-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl, phenylC₀-C₄alkyl,        pyridylC₀-C₄alkyl, pyrimidinylC₀-C₄alkyl, thienylC₀-C₄alkyl,        imidazolylC₀-C₄alkyl, pyrrolylC₀-C₄alkyl, pyrazolylC₀-C₄alkyl,        benzoisothiazolyl or tetrahydronapthyl, each of which is        substituted with from 0 to 4 substituents independently chosen        from R_(x), C₂-C₄alkanoyl, mono- and        di-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- and        di-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-membered        heterocycloalkyl)C₀-C₄alkyl and XR_(y); or    -   (ii) joined to R₅ to form, with the nitrogen to which R₄ and R₅        are bound, a heterocycle having from 1 to 3 rings, 5 to 7 ring        members in each ring, and is substituted with from 0 to 4        substituents independently chosen from R_(x), oxo and W—Z; and

-   R₅ is:    -   (i) hydrogen;    -   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,        (C₃-C₇carbocycle)C₀-C₄alkyl, each of which is substituted with        from 0 to 3 substituents independently chosen from halogen,        hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy, methylamino,        dimethylamino, trifluoromethyl and trifluoromethoxy; or    -   (iii) joined to R₄ to form an optionally substituted        heterocycle.

In certain other preferred compounds of Formula II, Ar is mono-, di-, ortri-substituted phenyl, optionally substituted naphthyl, or optionallysubstituted heteroaryl. In certain other preferred compounds of FormulaII, Ar is mono-, di-, or tri-substituted phenyl, or Ar is 1-naphthyl,2-naphthyl, pyridyl, pyrimidinyl, pyrazinyl, pyridizinyl, thienyl,thiazolyl, pyrazolyl, imidazolyl, tetrazolyl, oxazolyl, isoxazolyl,indazolyl, indolyl, pyrrolyl, furanyl, or triazolyl, each of which isoptionally mono-, di-, or tri-substituted.

In yet other preferred compounds of Formula II, Ar is phenyl substitutedwith between 1 and 3 residues independently selected from the groupconsisting of optionally substituted C₁₋₆alkyl, optionally substitutedC₂₋₆alkenyl, optionally substituted C₂₋₆alkynyl, optionally substitutedC₁₋₆alkoxy, optionally substituted (C₁₋₆alkoxy)C₁₋₆alkyl, optionallysubstituted (amino)C₁₋₆alkyl, optionally substituted mono- anddi-(C₁₋₆alkyl)amino.

Certain preferred compounds of Formula II include those compounds inwhich A is NR₄R₅ and are referred to herein as compounds of FormulaII-a. Certain preferred compounds of Formula II-a include thosecompounds in which: R₄ is chosen from (C₃-C₇cycloalkyl)C₀-C₄alkyl,phenylC₀-C₄alkyl, pyridylC₀-C₄alkyl, pyrimidinylC₀-C₄alkyl,thienylC₀-C₄alkyl, imidazolylC₀-C₄alkyl, pyrrolylC₀-C₄alkyl,pyrazolylC₀-C₄alkyl, indolylC₀-C₄alkyl, indazolylC₀-C₄alkyl,benzocycloalkenylC₀-C₄alkyl, decahydronaphthylC₀-C₄alkyl,benzoisothiazolylC₀-C₄alkyl, tetrahydroquinolinylC₀-C₄alkyl andtetrahydronaphthylC₀-C₄alkyl, each of which is substituted with from 0to 4 groups independently chosen from R_(x), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl, C₂-C₄alkanoyl and C₂-C₄alkanoyloxy; and R₅is C₁-C₆alkyl, C₂-C₆alkenyl or (C₃-C₇-carbocycle)C₀-C₄alkyl.

In other preferred compounds of Formula II-a provided herein includethose compounds in which R₄ and R₅ are joined to form a form a saturatedor partially saturated heterocycle containing 1 or 2 fused or spirorings; wherein the heterocycle is substituted with from 0 to 4substituents independently chosen from halogen, hydroxy, amino, cyano,—COOH, —CH₂COOH, —CO₂—C₁₋₆alkyl, —CH₂CO₂—C₁₋₆alkyl, —C(═O)NH₂,C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, mono- and di-(C₁-C₆alkyl)amino,C₁-C₆alkoxy, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,(C₃-C₇cycloalkyl)C₀-C₄alkyl, —S(O_(n))C₁-C₆alkyl, SO₃H, and phenyl. Morepreferably, R₄ and R₅ are joined to form a saturated 4- to 7-memberedheterocyclic ring that is substituted with from 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, C₁-C₂alkyl,C₁-C₂alkoxy, trifluoromethyl, difluoromethyl, trifluoromethoxydifluoromethoxy, —COOH, —CH₂COOH, —CO₂—C₁₋₂alkyl, and —CH₂CO₂—C₁₋₂alkyl.Certain preferred compounds of Formula II-a include those in which R₄and R₅ are combined to form an azepanyl, morpholinyl, homomorpholinyl,pyrrolidinyl, piperazinyl, homopiperazinyl, piperidinyl,homopiperidinyl, and the like.

In certain other preferred compounds of Formula II-a in which R₄ and R₅are combined to form a heterocycle, the heterocycle comprises 2 rings;wherein each of the rings is substituted with from 0 to 3 substituentsindependently selected from the group consisting of halogen, hydroxy,amino, cyano, C₁-C₂alkyl, C₁-C₂alkoxy, trifluoromethyl, difluoromethyl,trifluoromethoxy, and difluoromethoxy. Certain preferred compounds ofFormula II-a in which R₄ and R₅ are combined to form a bicyclicheterocycle include those in which the heterocycle istetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl,decahydroisoquinolinyl, indazolyl, indolinyl, phenylimidazolyl,pyridooxazinyl, benzoxazinyl, or the like.

The invention provides compounds Formula II and pharmaceutical saltsthereof, wherein the compound is according to Formula III:

wherein:

-   R₃ and R_(3a) are independently selected from the group consisting    of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy,    COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino;-   R₁₃ represents from 0 to 3 substituents independently chosen from:    -   (i) R_(x); and    -   (ii) phenyl and pyridyl, each of which is substituted with from        0 to 4 substituents independently chosen from halogen, hydroxy,        amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy,        (C₃-C₇cycloalkyl)C₀-C₄alkyl, C₁-C₂haloalkyl, C₁-C₂haloalkoxy and        mono- and di-(C₁-C₄alkyl)amino; and-   G is CH₂, sulfur, oxygen or NR_(E); wherein R_(E) is:    -   (i) hydrogen; or    -   (ii) C₁-C₆alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, phenyl or a 5- or        6-membered heteroaryl ring, each of which is substituted with        from 0 to 3 substituents independently chosen from R_(x). In        certain preferred compounds of Formula III, G is oxygen.

In other preferred compounds of Formula III, R₁₃ represents from 0 to 2substituents independently chosen from halogen, methyl, methoxy, ethyl,phenyl, and phenoxy, wherein each phenyl or phenoxy group is substitutedwith between 0 and 3 substituents chosen from R_(x).

Certain preferred compounds of Formula II-a include compounds orpharmaceutically acceptable forms thereof according to Formula (IV):

wherein:

-   R₃ and R_(3a) are independently selected from the group consisting    of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy,    COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino;-   R₁₀ and R₁₁ are independently chosen from hydrogen, C₁-C₆alkyl,    C₁-C₂haloalkyl and C₃-C₇cycloalkyl(C₀-C₂alkyl); and-   R₁₂ represents from 0 to 3 substituents independently chosen from    R_(x), mono- and di-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- and    di-(C₁-C₄alkyl)amino(C₁-C₄alkoxy) and YZ; or two adjacent R₁₂ groups    are joined to form a fused 5- to 7-membered carbocyclic or    heterocyclic ring.

The invention provides certain preferred compounds of Formula IV includethose compounds in which R₁₂ represents from 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, C₁-C₄alkyl,mono- and di-(C₁-C₂alkyl)amino, C₁-C₄alkoxy, C₁-C₂haloalkyl,C₁-C₂haloalkoxy and (C₃-C₇cycloalkyl)C₀-C₂alkyl.

Other preferred compounds of Formula IV include those compounds inwhich:

-   R₁ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy,    C₁-C₆haloalkyl, C₁-C₆haloalkoxy, (C₃-C₇cycloalkyl)-C₀-C₄alkyl;-   R₈ and R₉ are independently chosen from hydrogen, halogen, hydroxy,    C₁-C₆alkyl, C₁-C₆alkenyl, (C₃-C₆cycloalkyl)C₀-C₄alkyl and    C₁-C₆alkoxy; and-   Ar is phenyl, 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl,    pyrazinyl, pyridizinyl, thienyl, thiazolyl, pyrazolyl, imidazolyl,    tetrazolyl, oxazolyl, isoxazolyl, indazolyl, indolyl, pyrrolyl,    furanyl, and triazolyl, each of which is optionally mono-, di-, or    tri-substituted.

Other preferred compounds of Formula II-a include compounds orpharmaceutically acceptable forms thereof according to Formula (V):

wherein:

-   R₃ and R_(3a) are independently selected from the group consisting    of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy,    COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino;-   R₁₂ and R₁₃ independently represent from 0 to 3 substituents    independently chosen from R_(x);-   R₁₄ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,    C₁-C₂haloalkyl or (C₃-C₇cycloalkyl)C₀-C₂alkyl, COOH, CONH₂, CH₂COOH,    CH₂CONH₂, CO₂—C₁₋₆alkyl, CH₂CO₂—C₁₋₆alkyl, or SO₃H; and-   x is 0, 1 or 2, or in certain preferred compounds x is 1.

Certain preferred compounds of Formula V include those compounds inwhich R₁₂ and R₁₃ independently represent from 0 to 2 substituentsindependently chosen from halogen, methyl, methoxy and ethyl; and R₁₄ ishydrogen, C₁-C₆alkyl, C₂-C₆alkenyl or C₃-C₇cycloalkyl(C₀-C₂alkyl).

The invention provides certain preferred compounds of Formula V includethose compounds in which:

-   R₁ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy,    C₁-C₆haloalkyl, C₁-C₆haloalkoxy, (C₃-C₇cycloalkyl)-C₀-C₄alkyl;-   R₈ and R₉ are independently chosen from hydrogen, halogen, hydroxy,    C₁-C₆alkenyl, (C₃-C₆cycloalkyl)C₀-C₄alkyl and C₁-C₆alkoxy; and-   Ar is phenyl which is mono-, di-, or tri-substituted, or 1-naphthyl,    2-naphthyl, pyridyl, pyrimidinyl, pyrazinyl, pyridizinyl, thienyl,    thiazolyl, pyrazolyl, imidazolyl, tetrazolyl, oxazolyl, isoxazolyl,    pyrrolyl, furanyl, indolyl, indazolyl, and triazolyl, each of which    is optionally mono-, di-, or tri-substituted.

Yet other preferred compounds of Formula II-a include compounds orpharmaceutically acceptable forms thereof according to Formula (VI):

wherein:

-   R₃ and R_(3a) are independently selected from the group consisting    of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy,    COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino;-   R₁₂ and R₁₃ represent from 0 to 3 substituents independently chosen    from R_(x);-   G is CH₂, NH, sulfur or oxygen;-   G₃ is N, CH, or CR_(x) and-   x is 0, 1 or 2, or in certain preferred compounds x is 1.

Other preferred compounds of Formula II-a provided herein include thosecompounds which satisfy Formula VII

wherein

-   R₃ and R_(3a) are independently selected from the group consisting    of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy,    COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino;-   R₁₂ and R₁₃ independently represent from 0 to 3 substituents    independently chosen from R_(x);-   G is CH₂, NH or oxygen; and-   x is 0, 1 or 2, or in certain preferred compounds x is 1.

In certain preferred compounds of Formula VI or Formula VII provided bythe invention, R₁₂ and R₁₃ independently represent from 0 to 3substituents independently chosen from halogen, hydroxy, amino, cyano,C₁-C₄alkyl, mono- and di-(C₁-C₂alkyl)amino, C₁-C₄alkoxy, C₁-C₂haloalkyl,C₁-C₂haloalkoxy, and (C₃-C₇cycloalkyl)C₀-C₂alkyl. Other preferredcompounds of Formula VI and Formula VII include those in which R₁₂ andR₁₃ independently represent from 0 to 2 substituents independentlychosen from halogen, methyl, methoxy and ethyl.

Other preferred compounds of Formula VII include those in which R₅ isC₁-C₆alkyl; and R₁₂ and R₁₃ each represent from 0 to 2 substituentsindependently chosen from halogen, methyl, methoxy and ethyl.

Other preferred compounds of Formula II provided herein include thosecompounds, which are herein defined as compounds of Formula II-b, inwhich:

-   A is OR₄; and-   R₄ is C₂-C₆alkyl, C₂-C₆alkenyl, phenylC₀-C₄alkyl,    naphthylC₀-C₄alkyl, pyridylC₀-C₄alkyl, pyrimidinylC₀-C₄alkyl,    thienylC₀-C₄alkyl, imidazolylC₀-C₄alkyl or pyrrolylC₀-C₄alkyl, each    of which is substituted with from 0 to 4 substituents independently    chosen from R_(x), mono- and di-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono-    and di-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-membered    heterocycloalkyl)C₀-C₄alkyl and C₂-C₄alkanoyl.

Certain preferred compounds of Formula II-b include those compounds inwhich R₄ is phenyl, benzyl, pyridyl or pyridylmethyl, each of which issubstituted with from 0 to 4 substituents independently chosen fromR_(x), mono- and di-C₁-C₄alkylamino(C₀-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl and C₂-C₄alkanoyl.

Yet other preferred compounds provided herein include those compounds ofFormula II-b which satisfy Formula VIII:

wherein:

-   D is CH or N;-   R₃ and R_(3a) are independently selected from the group consisting    of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy,    COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino;-   R₂₁ represents from 0 to 3 substituents independently chosen from    R_(x) and LR_(d); or two adjacent R₂₁ groups are joined to form a    fused 5- to 7-membered carbocyclic or heterocyclic ring that is    substituted with from 0 to 3 substituents independently chosen from    R_(x);-   L is a single bond or —CH₂—; and-   R_(d) is piperazinyl, morpholinyl, piperidinyl or pyrrolidinyl.

Certain preferred compounds according to Formula VIII provided hereininclude those compounds in which:

-   R₂₁ represents from 0 to 3 substituents independently chosen from    R_(x) and LR_(d);-   R₁ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy,    C₁-C₆haloalkyl, C₁-C₆haloalkoxy, (C₃-C₇cycloalkyl)-C₀-C₄alkyl;-   R₈ and R₉ are independently chosen from hydrogen, halogen, hydroxy,    C₁-C₆alkyl, C₁-C₆alkenyl, (C₃-C₆cycloalkyl)C₀-C₄alkyl and    C₁-C₆alkoxy; and-   Ar is phenyl which is mono-, di-, or tri-substituted, or 1-naphthyl,    2-naphthyl, pyridyl, pyrimidinyl, pyrazinyl, pyridizinyl, thienyl,    thiazolyl, pyrazolyl, imidazolyl, tetrazolyl, oxazolyl, isoxazolyl,    pyrrolyl, furanyl, indolyl, indazolyl, and triazolyl, each of which    is optionally mono-, di-, or tri-substituted.

Yet other preferred compounds of Formula VIII include those compounds inwhich the group designated:

is chosen from naphthyl, tetrahydronaphthyl, benzofuranyl,benzodioxolyl, indanyl, indolyl, indazolyl, benzodioxolyl,benzo[1,4]dioxanyl and benzoxazolyl, each of which is substituted withfrom 0 to 3 substituents independently chosen from R_(x).

Certain preferred compounds of any one of Formula II, II-a, II-b, III,IV, V, VI, VII, or VIII include those compounds in which the Arsubstituent is mono-, di-, or tri-substituted phenyl, optionallysubstituted naphthyl, or optionally substituted heteroaryl. In otherpreferred compounds of any one of Formula II, II-a, II-b, III, IV, V,VI, VII, or VIII, Ar is selected from the group consisting of mono-,di-, or tri-substituted phenyl, or Ar is 1-naphthyl, 2-naphthyl,pyridyl, pyrimidinyl, pyrazinyl, pyridizinyl, thienyl, thiazolyl,pyrazolyl, imidazolyl, tetrazolyl, oxazolyl, isoxazolyl, indazolyl,indolyl, pyrrolyl, furanyl, and triazolyl, each of which is optionallymono-, di-, or tri-substituted

Certain other preferred compounds of any one of Formula II, II-a, II-b,III, IV, V, VI, VII, or VIII include those compounds in which the Arsubstituent is phenyl substituted with between 1 and 3 residuesindependently selected from the group consisting of optionallysubstituted C₁₋₆alkyl, optionally substituted C₂₋₆alkenyl, optionallysubstituted C₂₋₆alkynyl, optionally substituted C₁₋₆alkoxy, optionallysubstituted (C₁₋₆alkoxy)C₁₋₆alkyl, optionally substituted(amino)C₁₋₆alkyl, optionally substituted mono- and di-(C₁₋₆alkyl)amino.

Certain preferred compounds of Formula II, II-a, II-b, III, IV, V, VI,VII, or VIII include those compounds in which R₁ is hydrogen,C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy, C₁-C₆haloalkyl,C₁-C₆haloalkoxy, (C₃-C₇cycloalkyl)-C₀-C₄alkyl. In other preferredcompounds of Formula II, II-a, II-b, III, IV, V, VI, VII, or VIII, R₁ ishydrogen, C₁-C₄alkyl or C₁-C₄alkoxy, or more preferably R₁ is hydrogen,methyl, ethyl, or methoxy.

Yet other preferred compounds of Formula II, II-a, II-b, III, IV, V, VI,VII, or VIII include those compounds in which R₃ represents between 0and 2 substituents, each of which is independently selected fromC₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy, hydroxy, COOH,CONH₂, SO₂NH₂, mono- and di-(C₁₋₆alkyl)amino, (amino)C₀₋₆alkyl. Incertain other preferred compounds of Formula II, II-a, II-b, III, IV, V,VI, VII, or VIII, R₃ represents between 0 and 2 substituents, each ofwhich is independently selected from C₁₋₆alkyl, C₁₋₆alkoxy, hydroxy,COOH, CONH₂, and SO₂NH₂.

Certain preferred compounds of Formula IX, include those compounds inwhich Ar is 1-naphthyl, 2-naphthyl, pyridyl, pyrimidinyl, pyrazinyl,pyridizinyl, thienyl, thiazolyl, pyrazolyl, imidazolyl, tetrazolyl,oxazolyl, isoxazolyl, indazolyl, indolyl, benzoimidazolyl, pyrrolyl,furanyl, or triazolyl, each of which is optionally mono-, di-, ortri-substituted. More preferably, the Ar substituent is unsubstituted orsubstituted with between one and three groups independently selectedfrom optionally substituted C₁₋₆alkyl, optionally substitutedC₂₋₆alkenyl, optionally substituted C₂₋₆alkynyl, optionally substitutedC₁₋₆alkoxy, optionally substituted (C₁₋₆alkoxy)C₁₋₆alkyl, optionallysubstituted (amino)C₁₋₆alkyl, or optionally substituted mono- anddi-(C₁₋₆alkyl)amino.

In certain other preferred compounds according to Formula IX, Ar isindazolyl, indolyl, or benzoimidazolyl, each of which is optionallysubstituted with between 1 and 3 residues independently selected fromthe group consisting of optionally substituted C₁₋₆alkyl, optionallysubstituted C₂₋₆alkenyl, optionally substituted C₂₋₆alkynyl, optionallysubstituted C₁₋₆alkoxy, optionally substituted (C₁₋₆alkoxy)C₁₋₆alkyl,optionally substituted (amino)C₁₋₆alkyl, optionally substituted mono-and di-(C₁₋₆alkyl)amino.

Other preferred compounds according to Formula IX include thosecompounds in which

-   R₁ is hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, halogen, C₁₋₆haloalkyl,    C₁₋₆haloalkoxy, C₃₋₈cycloalkyl, C₃₋₈cycloalkyl-C₁₋₆alkyl; and-   R₃ represents between 0 and 2 substituents, each of which is    independently selected from C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,    C₁₋₆haloalkoxy, mono- and di-(C₁₋₆alkyl)amino, (amino)C₀₋₆alkyl.

Yet other preferred compounds according to Formula IX include thosecompounds in which

-   R₄ is chosen from (C₃-C₇cycloalkyl)C₀-C₄alkyl, phenylC₀-C₄alkyl,    pyridylC₀-C₄alkyl, pyrimidinylC₀-C₄alkyl, thienylC₀-C₄alkyl,    imidazolylC₀-C₄alkyl, pyrrolylC₀-C₄alkyl, pyrazolylC₀-C₄alkyl,    indolylC₀-C₄alkyl, indazolylC₀-C₄alkyl, benzocycloalkenylC₀-C₄alkyl,    decahydronaphthylC₀-C₄alkyl, benzoisothiazolylC₀-C₄alkyl,    tetrahydroquinolinylC₀-C₄alkyl and tetrahydronaphthylC₀-C₄alkyl,    each of which is substituted with from 0 to 4 groups independently    chosen from R_(x), mono- and di-C₁-C₄alkylamino(C₁-C₄alkyl), mono-    and di-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-membered    heterocycloalkyl)C₀-C₄alkyl, C₂-C₄alkanoyl and C₂-C₄alkanoyloxy; and-   R₅ is C₁-C₆alkyl, C₂-C₆alkenyl or (C₃-C₇carbocycle)C₀-C₄alkyl.

In certain other compounds of Formula IX, R₄ and R₅ are joined to form asaturated or partially saturated heterocycle containing 1 or 2 fused orspiro rings; wherein the heterocycle is substituted with from 0 to 4substituents independently chosen from halogen, hydroxy, amino, cyano,—COOH, —CH₂COOH, —CO₂—C₁₋₆alkyl, —CH₂CO₂—C₁₋₆alkyl, C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, mono- and di-(C₁-C₆alkyl)amino, C₁-C₆alkoxy,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, (C₃-C₇cycloalkyl)C₀-C₄alkyl,—S(O_(n))C₁-C₆alkyl, SO₃H, and phenyl. In certain other compounds ofFormula IX, R₄ and R₅ are joined to form a saturated 4- to 7-memberedheterocyclic ring that is substituted with from 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, C₁-C₂alkoxy,trifluoromethyl, difluoromethyl, trifluoromethoxy difluoromethoxy,—COOH, —CH₂COOH, —CO₂—C₁₋₂alkyl, and —CH₂CO₂—C₁₋₂alkyl. In certainparticularly preferred compounds of Formula IX, the heterocyclic ringformed by the joining of R₄ and R₅ is azepanyl, morpholinyl,homomorpholinyl, pyrrolidinyl, piperazinyl, homopiperazinyl,piperidinyl, or homopiperidinyl.

Certain compounds according to the Formulas provided herein, which havetwo or more stereogenic centers, have a diastereomeric excess of atleast 50%. For example, such compounds may have a diastereomeric excessof at least 60%, 70%, 80%, 85%, 90%, 95%, or 98%. Certain such compoundshave a diastereomeric excess of at least 99%.

Certain compounds according to the Formulas provided herein, which haveone or more stereogenic center, have an enantiomeric excess of at least50%. For example, such compounds may have an enantiomeric excess of atleast 60%, 70%, 80%, 85%, 90%, 95%, or 98%. Certain such compounds havean enantiomeric excess of at least 99%. It will be apparent that singleenantiomers (optically active forms) can be obtained by asymmetricsynthesis, synthesis from optically pure precursors or by resolution ofthe racemates. Resolution of the racemates can be accomplished, forexample, by conventional methods such as crystallization in the presenceof a resolving agent, or chromatography, using, for example a chiralHPLC column

1-Aryl-4-substituted isoquinolines and pharmaceutically acceptable formsthereof provided herein detectably alter (modulate) C5a receptoractivity and/or ligand binding, as determined using a standard in vitroC5 receptor-mediated chemotaxis assay (described in Example 55),radioligand binding (described in Example 60), or C5a receptor-mediatedcalcium mobilization assay (described in Example 62). Preferredcompounds exhibit an EC₅₀ of about 500 nM or less in such a standard C5areceptor-mediated chemotaxis, radioligand binding, and/or calciummobilization assay, more preferably an EC₅₀ of about 250 nM or less insuch an assay, still more preferably an EC₅₀ of about 200, 150, 100, 50,25, 10, or 5 nM or less in such an assay.

Initial characterization of compounds can be conveniently carried outusing a C5a receptor binding assay or functional assay, such as setforth in the Examples, and may be expedited by applying such assays in ahigh throughput screening setting. Additional assays suitable fordetermining the effects of small molecule compounds on C5a receptorbinding and receptor modulatory activity, as well as assays suitable formeasuring their effects on C5a-induced neutropenia in vivo, can be foundin the published literature, for example in U.S. Pat. No. 5,807,824,which is incorporated herein by reference for its disclosure in thisregard in Examples 6-9, columns 19-23, as well as for its discussion ofcomplement and inflammation at columns 1-2. Those of skill in the artwill recognize that such assays can be readily adapted to the use ofcells or animals of different species as deemed appropriate.

In certain embodiments, preferred compounds have favorablepharmacological properties, including oral bioavailability (such that asub-lethal or preferably a pharmaceutically acceptable oral dose,preferably less than 2 grams, more preferably of less than or equal toone gram, can provide a detectable in vivo effect such as a reduction ofC5a-induced neutropenia), ability to inhibit leukocyte chemotaxis atnanomolar concentrations and preferably at sub-nanomolar concentrations,low toxicity (a preferred compound is nontoxic when a C5areceptor-modulatory amount is administered to a subject), minimal sideeffects (a preferred compound produces side effects comparable toplacebo when a C5a receptor-modulatory amount of the compound isadministered to a subject), low serum protein binding, and a suitable invitro and in vivo half-life (a preferred compound exhibits an in vitrohalf-life that is equal to an in vivo half-life allowing for Q.I.D.dosing, preferably T.I.D. dosing, more preferably B.I.D. dosing, andmost preferably once-a-day dosing). Distribution in the body to sites ofcomplement activity is also desirable (e.g., compounds used to treat CNSdisorders will preferably penetrate the blood brain barrier, while lowbrain levels of compounds used to treat peripheral disorders aretypically preferred).

Routine assays that are well known in the art may be used to assessthese properties, and identify superior compounds for a particular use.For example, assays used to predict bioavailability include transportacross human intestinal cell monolayers, such as Caco-2 cell monolayers.Penetration of the blood brain barrier of a compound in humans may bepredicted from the brain levels of the compound in laboratory animalsgiven the compound (e.g., intravenously). Serum protein binding may bepredicted from albumin binding assays, such as those described byOravcová, et al. (1996) Journal of Chromatography B 677:1-27. Compoundhalf-life is inversely proportional to the frequency of dosage of acompound required to achieve an C5a receptor modulatory amount. In vitrohalf-lives of compounds may be predicted from assays of microsomalhalf-life as described by Kuhnz and Gieschen (1998) Drug Metabolism andDisposition 26:1120-27.

As noted above, preferred compounds provided herein are nontoxic. Ingeneral, the term “nontoxic” as used herein shall be understood in arelative sense and is intended to refer to any substance that has beenapproved by the United States Food and Drug Administration (“FDA”) foradministration to mammals (preferably humans) or, in keeping withestablished criteria, is susceptible to approval by the FDA foradministration to mammals (preferably humans). In addition, a highlypreferred nontoxic compound generally satisfies one or more of thefollowing criteria: (1) does not substantially inhibit cellular ATPproduction; (2) does not significantly prolong heart QT intervals; (3)does not cause substantial liver enlargement, and (4) does not causesubstantial release of liver enzymes.

As used herein, a compound that “does not substantially inhibit cellularATP production” is a compound that satisfies the criteria set forth inExample 64, herein. In other words, cells treated as described inExample 64 with 100 μM of such a compound exhibit ATP levels that are atleast 50% of the ATP levels detected in untreated cells. In more highlypreferred embodiments, such cells exhibit ATP levels that are at least80% of the ATP levels detected in untreated cells.

A compound that “does not significantly prolong heart QT intervals” is acompound that does not result in a statistically significantprolongation of heart QT intervals (as determined byelectrocardiography) in guinea pigs, minipigs or dogs uponadministration of twice the minimum dose yielding a therapeuticallyeffective in vivo concentration. In certain preferred embodiments, adose of 0.01, 0.05. 0.1, 0.5, 1, 5, 10, 40 or 50 mg/kg administeredparenterally or orally does not result in a statistically significantprolongation of heart QT intervals. By “statistically significant” ismeant results varying from control at the p<0.1 level or more preferablyat the p<0.05 level of significance as measured using a standardparametric assay of statistical significance such as a student's T test.

A compound “does not cause substantial liver enlargement” if dailytreatment of laboratory rodents (e.g., mice or rats) for 5-10 days withtwice the minimum dose that yields a therapeutically effective in vivoconcentration results in an increase in liver to body weight ratio thatis no more than 100% over matched controls. In more highly preferredembodiments, such doses do not cause liver enlargement of more than 75%or 50% over matched controls. If non-rodent mammals (e.g., dogs) areused, such doses should not result in an increase of liver to bodyweight ratio of more than 50%, preferably not more than 25%, and morepreferably not more than 10% over matched untreated controls. Preferreddoses within such assays include 0.01, 0.05. 0.1, 0.5, 1, 5, 10, 40 or50 mg/kg administered parenterally or orally.

Similarly, a compound “does not promote substantial release of liverenzymes” if administration of twice the minimum dose yielding atherapeutically effective in vivo concentration does not elevate serumlevels of ALT, LDH or AST in laboratory rodents by more than 100% overmatched mock-treated controls. In more highly preferred embodiments,such doses do not elevate such serum levels by more than 75% or 50% overmatched controls. Alternately, a compound “does not promote substantialrelease of liver enzymes” if, in an in vitro hepatocyte assay,concentrations (in culture media or other such solutions that arecontacted and incubated with hepatocytes in vitro) equivalent totwo-fold the minimum in vivo therapeutic concentration of the compounddo not cause detectable release of any of such liver enzymes intoculture medium above baseline levels seen in media from matchedmock-treated control cells. In more highly preferred embodiments, thereis no detectable release of any of such liver enzymes into culturemedium above baseline levels when such compound concentrations arefive-fold, and preferably ten-fold the minimum in vivo therapeuticconcentration of the compound.

In other embodiments, certain preferred compounds do not inhibit orinduce microsomal cytochrome P450 enzyme activities, such as CYP1A2activity, CYP2A6 activity, CYP2C9 activity, CYP2C19 activity, CYP2D6activity, CYP2E1 activity or CYP3A4 activity at a concentration equal tothe minimum therapeutically effective in vivo concentration.

Certain preferred compounds are not clastogenic or mutagenic (e.g., asdetermined using standard assays such as the Chinese hamster ovary cellvitro micronucleus assay, the mouse lymphoma assay, the human lymphocytechromosomal aberration assay, the rodent bone marrow micronucleus assay,the Ames test or the like) at a concentration equal to the minimumtherapeutically effective in vivo concentration. In other embodiments,certain preferred compounds do not induce sister chromatid exchange(e.g., in Chinese hamster ovary cells) at such concentrations.

In certain embodiments, preferred compounds exert theirreceptor-modulatory effects with high specificity. This means that theyonly bind to, activate, or inhibit the activity of certain receptorsother than C5a receptors with affinity constants of greater than 100nanomolar, preferably greater than 1 micromolar, more preferably greaterthan 4 micromolar. Also provided herein are highly specific C5a receptormodulatory compounds that exhibit 200-fold greater affinity for the C5areceptor that for other cellular receptors. Such receptors includeneurotransmitter receptors such as alpha- or beta-adrenergic receptors,muscarinic receptors (particularly m1, m2 or m3 receptors), dopaminereceptors, and metabotropic glutamate receptors; as well as histaminereceptors and cytokine receptors (e.g., interleukin receptors,particularly IL-8 receptors). Such receptors may also include GABA_(A)receptors, bioactive peptide receptors (other than C5a receptors and C3areceptors, including NPY or VIP receptors), neurokinin receptors,bradykinin receptors, and hormone receptors (e.g., CRF receptors,thyrotropin releasing hormone receptors or melanin-concentrating hormonereceptors). Compounds that act with high specificity generally exhibitfewer undesirable side effects.

Within certain embodiments, modulators provided herein do not binddetectably to receptors that do not mediate inflammatory responses, suchas GABA receptors, MCH receptors, NPY receptors, dopamine receptors,serotonin receptors and VR1 receptors, with high or even moderateaffinity. In addition, or alternatively, certain preferred C5a receptormodulators exhibit an affinity for C5a receptor that is substantiallyhigher than for receptors that do not mediate inflammatory responses(e.g., at least five times higher, at least ten times higher or at least100 times higher). Assays for evaluating binding to receptors that donot mediate inflammatory responses include, for example, those describedin U.S. Pat. No. 6,310,212, which is incorporated herein by referencefor its disclosure of a GABA_(A) receptor binding assays in Examples 14,columns 16-17, in U.S. patent application Ser. No. 10/152,189 which isincorporated herein by reference for its disclosure of an MCH receptorbinding assay in Example 2, pages 104-105, in U.S. Pat. No. 6,362,186,which is incorporated herein by reference for its disclosure of CRF, andNPY receptor binding assays in Examples 19, columns 45-46, in U.S. Pat.No. 6,355,644, which is incorporated herein by reference for itsdisclosure of a dopamine receptor binding assay at column 10, and inU.S. Pat. No. 6,482,611, which is incorporated herein by reference forits disclosure of VR1 receptor binding assays in Examples 4-5, column14. It will be apparent that the C5a receptor modulators provided hereinmay, but need not, bind to one or more other receptors known to mediateinflammatory responses, such as C3a receptors and/or A₃ receptors.

Certain preferred compounds are C5a receptor antagonists that do notpossess significant (e.g., greater than 5%) agonist activity in any ofthe C5a receptor-mediated functional assays discussed herein.Specifically, this undesired agonist activity can be evaluated, forexample, in the GTP binding assay of Example 61, by measuring smallmolecule mediated GTP binding in the absence of the natural agonist,C5a. Similarly, in a calcium mobilization assay (e.g., that of Example62) a small molecule compound can be directly assayed for the ability ofthe compound to stimulate calcium levels in the absence of the naturalagonist, C5a. The preferred extent of C5a agonist activity exhibited bycompounds provided herein is less than 10%, 5% or 2% of the responseelicited by the natural agonist, C5a.

Also preferred, in certain embodiments, are C5a receptor modulators thatinhibit the occurrence of C5a-induced oxidative burst (OB) ininflammatory cells (e.g., neutrophil) as can be conveniently determinedusing an in vitro neutrophil OB assay.

For detection purposes, compounds provided herein may beisotopically-labeled or radiolabeled. Accordingly, compounds recited inFormula I (or any other formula specifically recited herein) may haveone or more atoms replaced by an atom of the same element having anatomic mass or mass number different from the atomic mass or mass numberusually found in nature. Examples of isotopes that can be present incompounds provided herein include isotopes of hydrogen, carbon,nitrogen, oxygen, phosphorous, fluorine and chlorine, such as ²H, ³H,¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F and ³⁶Cl. In addition,substitution with heavy isotopes such as deuterium (i.e., ²H) can affordcertain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements and, hence, may be preferred in some circumstances.

Pharmaceutical Compositions

The present invention also provides pharmaceutical compositionscomprising one or more C5a receptor modulators provided herein, togetherwith at least one physiologically acceptable carrier or excipient.Pharmaceutical compositions may comprise, for example, one or more ofwater, buffers (e.g., neutral buffered saline or phosphate bufferedsaline), ethanol, mineral oil, vegetable oil, dimethylsulfoxide,carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol,proteins, adjuvants, polypeptides or amino acids such as glycine,antioxidants, chelating agents such as EDTA or glutathione and/orpreservatives. As noted above, other active ingredients may (but neednot) be included in the pharmaceutical compositions provided herein.

Pharmaceutical compositions may be formulated for any appropriate mannerof administration, including, for example, topical, oral, nasal, rectalor parenteral administration. The term parenteral as used hereinincludes subcutaneous, intradermal, intravascular (e.g., intravenous),intramuscular, spinal, intracranial, intrathecal and intraperitonealinjection, as well as any similar injection or infusion technique. Incertain embodiments, compositions in a form suitable for oral use arepreferred. Such forms include, for example, tablets, troches, lozenges,aqueous or oily suspensions, dispersible powders or granules, emulsion,hard or soft capsules, or syrups or elixirs. Within yet otherembodiments, compositions provided herein may be formulated as alyophilizate. Formulation for topical administration may be preferredfor certain conditions (e.g., in the treatment of skin conditions suchas burns or itch).

Compositions intended for oral use may further comprise one or morecomponents such as sweetening agents, flavoring agents, coloring agentsand/or preserving agents in order to provide appealing and palatablepreparations. Tablets contain the active ingredient in admixture withphysiologically acceptable excipients that are suitable for themanufacture of tablets. Such excipients include, for example, inertdiluents (e.g., calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate), granulating and disintegrating agents(e.g., corn starch or alginic acid), binding agents (e.g., starch,gelatin or acacia) and lubricating agents (e.g., magnesium stearate,stearic acid or talc). The tablets may be uncoated or they may be coatedby known techniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonosterate or glyceryl distearate may be employed.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent(e.g., calcium carbonate, calcium phosphate or kaolin), or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium (e.g., peanut oil, liquid paraffin or olive oil).

Aqueous suspensions contain the active material(s) in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients include suspending agents (e.g., sodiumcarboxymethylcellulose, methylcellulose, hydropropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia);and dispersing or wetting agents (e.g., naturally-occurring phosphatidessuch as lecithin, condensation products of an alkylene oxide with fattyacids such as polyoxyethylene stearate, condensation products ofethylene oxide with long chain aliphatic alcohols such asheptadecaethyleneoxycetanol, condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides such as polyethylene sorbitan monooleate). Aqueoussuspensions may also comprise one or more preservatives, for exampleethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, oneor more flavoring agents, and one or more sweetening agents, such assucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientsin a vegetable oil (e.g., arachis oil, olive oil, sesame oil or coconutoil) or in a mineral oil such as liquid paraffin. The oily suspensionsmay contain a thickening agent such as beeswax, hard paraffin or cetylalcohol. Sweetening agents such as those set forth above, and/orflavoring agents may be added to provide palatable oral preparations.Such suspensions may be preserved by the addition of an anti-oxidantsuch as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, such as sweetening, flavoring and coloringagents, may also be present.

Pharmaceutical compositions may also be in the form of oil-in-wateremulsions. The oily phase may be a vegetable oil (e.g., olive oil orarachis oil), a mineral oil (e.g., liquid paraffin) or a mixturethereof. Suitable emulsifying agents include naturally-occurring gums(e.g., gum acacia or gum tragacanth), naturally-occurring phosphatides(e.g., soy bean lecithin, and esters or partial esters derived fromfatty acids and hexitol), anhydrides (e.g., sorbitan monoleate) andcondensation products of partial esters derived from fatty acids andhexitol with ethylene oxide (e.g., polyoxyethylene sorbitan monoleate).An emulsion may also comprise one or more sweetening and/or flavoringagents.

Syrups and elixirs may be formulated with sweetening agents, such asglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso comprise one or more demulcents, preservatives, flavoring agentsand/or coloring agents.

Formulations for topical administration typically comprise a topicalvehicle combined with active agent(s), with or without additionaloptional components. Suitable topical vehicles and additional componentsare well known in the art, and it will be apparent that the choice of avehicle will depend on the particular physical form and mode ofdelivery. Topical vehicles include water; organic solvents such asalcohols (e.g., ethanol or isopropyl alcohol) or glycerin; glycols(e.g., butylene, isoprene or propylene glycol); aliphatic alcohols(e.g., lanolin); mixtures of water and organic solvents and mixtures oforganic solvents such as alcohol and glycerin; lipid-based materialssuch as fatty acids, acylglycerols (including oils, such as mineral oil,and fats of natural or synthetic origin), phosphoglycerides,sphingolipids and waxes; protein-based materials such as collagen andgelatin; silicone-based materials (both non-volatile and volatile); andhydrocarbon-based materials such as microsponges and polymer matrices. Acomposition may further include one or more components adapted toimprove the stability or effectiveness of the applied formulation, suchas stabilizing agents, suspending agents, emulsifying agents, viscosityadjusters, gelling agents, preservatives, antioxidants, skin penetrationenhancers, moisturizers and sustained release materials. Examples ofsuch components are described in Martindale—The Extra Pharmacopoeia(Pharmaceutical Press, London 1993) and Martin (ed.), Remington'sPharmaceutical Sciences. Formulations may comprise microcapsules, suchas hydroxymethylcellulose or gelatin-microcapsules, liposomes, albuminmicrospheres, microemulsions, nanoparticles or nanocapsules.

A topical formulation may be prepared in a variety of physical formsincluding, for example, solids, pastes, creams, foams, lotions, gels,powders, aqueous liquids and emulsions. The physical appearance andviscosity of such forms can be governed by the presence and amount ofemulsifier(s) and viscosity adjuster(s) present in the formulation.Solids are generally firm and non-pourable and commonly are formulatedas bars or sticks, or in particulate form; solids can be opaque ortransparent, and optionally can contain solvents, emulsifiers,moisturizers, emollients, fragrances, dyes/colorants, preservatives andother active ingredients that increase or enhance the efficacy of thefinal product. Creams and lotions are often similar to one another,differing mainly in their viscosity; both lotions and creams may beopaque, translucent or clear and often contain emulsifiers, solvents,and viscosity adjusting agents, as well as moisturizers, emollients,fragrances, dyes/colorants, preservatives and other active ingredientsthat increase or enhance the efficacy of the final product. Gels can beprepared with a range of viscosities, from thick or high viscosity tothin or low viscosity. These formulations, like those of lotions andcreams, may also contain solvents, emulsifiers, moisturizers,emollients, fragrances, dyes/colorants, preservatives and other activeingredients that increase or enhance the efficacy of the final product.Liquids are thinner than creams, lotions, or gels and often do notcontain emulsifiers. Liquid topical products often contain solvents,emulsifiers, moisturizers, emollients, fragrances, dyes/colorants,preservatives and other active ingredients that increase or enhance theefficacy of the final product.

Suitable emulsifiers for use in topical formulations include, but arenot limited to, ionic emulsifiers, cetearyl alcohol, non-ionicemulsifiers like polyoxyethylene oleyl ether, PEG-40 stearate,ceteareth-12, ceteareth-20, ceteareth-30, ceteareth alcohol, PEG-100stearate and glyceryl stearate. Suitable viscosity adjusting agentsinclude, but are not limited to, protective colloids or non-ionic gumssuch as hydroxyethylcellulose, xanthan gum, magnesium aluminum silicate,silica, microcrystalline wax, beeswax, paraffin, and cetyl palmitate. Agel composition may be formed by the addition of a gelling agent such aschitosan, methyl cellulose, ethyl cellulose, polyvinyl alcohol,polyquatemiums, hydroxyethylcellulose, hydroxypropylcellulose,hydroxypropylmethylcellulose, carbomer or ammoniated glycyrrhizinate.Suitable surfactants include, but are not limited to, nonionic,amphoteric, ionic and anionic surfactants. For example, one or more ofdimethicone copolyol, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 80, lauramide DEA, cocamide DEA, and cocamide MEA, oleylbetaine, cocamidopropyl phosphatidyl PG-dimonium chloride, and ammoniumlaureth sulfate may be used within topical formulations. Suitablepreservatives include, but are not limited to, antimicrobials such asmethylparaben, propylparaben, sorbic acid, benzoic acid, andformaldehyde, as well as physical stabilizers and antioxidants such asvitamin E, sodium ascorbate/ascorbic acid and propyl gallate. Suitablemoisturizers include, but are not limited to, lactic acid and otherhydroxy acids and their salts, glycerin, propylene glycol, and butyleneglycol. Suitable emollients include lanolin alcohol, lanolin, lanolinderivatives, cholesterol, petrolatum, isostearyl neopentanoate andmineral oils. Suitable fragrances and colors include, but are notlimited to, FD&C Red No. 40 and FD&C Yellow No. 5. Other suitableadditional ingredients that may be included a topical formulationinclude, but are not limited to, abrasives, absorbents, anti-cakingagents, anti-foaming agents, anti-static agents, astringents (e.g.,witch hazel, alcohol and herbal extracts such as chamomile extract),binders/excipients, buffering agents, chelating agents, film formingagents, conditioning agents, propellants, opacifying agents, pHadjusters and protectants.

An example of a suitable topical vehicle for formulation of a gel is:hydroxypropylcellulose (2.1%); 70/30 isopropyl alcohol/water (90.9%);propylene glycol (5.1%); and Polysorbate 80 (1.9%). An example of asuitable topical vehicle for formulation as a foam is: cetyl alcohol(1.1%); stearyl alcohol (0.5%; Quaternium 52 (1.0%); propylene glycol(2.0%); Ethanol 95 PGF3 (61.05%); deionized water (30.05%); P75hydrocarbon propellant (4.30%). All percents are by weight.

Typical modes of delivery for topical compositions include applicationusing the fingers; application using a physical applicator such as acloth, tissue, swab, stick or brush; spraying (including mist, aerosolor foam spraying); dropper application; sprinkling; soaking; andrinsing. Controlled release vehicles can also be used.

A pharmaceutical composition may be prepared as a sterile injectibleaqueous or oleaginous suspension. The modulator, depending on thevehicle and concentration used, can either be suspended or dissolved inthe vehicle. Such a composition may be formulated according to the knownart using suitable dispersing, wetting agents and/or suspending agentssuch as those mentioned above. Among the acceptable vehicles andsolvents that may be employed are water, 1,3-butanediol, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils may be employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed, including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use inthe preparation of injectible compositions, and adjuvants such as localanesthetics, preservatives and/or buffering agents can be dissolved inthe vehicle.

C5a modulators described herein may be formulated as inhaledformulations, including sprays, mists, or aerosols. Such formulationsare particularly useful for the treatment of asthma or other respiratoryconditions. For inhalation formulations, the compounds provided hereinmay be delivered via any inhalation methods known to those skilled inthe art. Such inhalation methods and devices include, but are notlimited to, metered dose inhalers with propellants such as CFC or FIFAor propellants that are physiologically and environmentally acceptable.Other suitable devices are breath operated inhalers, multidose drypowder inhalers and aerosol nebulizers. Aerosol formulations for use inthe subject method typically include propellants, surfactants andco-solvents and may be filled into conventional aerosol containers thatare closed by a suitable metering valve.

Inhalant compositions may comprise liquid or powdered compositionscontaining the active ingredient that are suitable for nebulization andintrabronchial use, or aerosol compositions administered via an aerosolunit dispensing metered doses. Suitable liquid compositions comprise theactive ingredient in an aqueous, pharmaceutically acceptable inhalantsolvent, e.g., isotonic saline or bacteriostatic water. The solutionsare administered by means of a pump or squeeze-actuated nebulized spraydispenser, or by any other conventional means for causing or enablingthe requisite dosage amount of the liquid composition to be inhaled intothe patient's lungs. Suitable formulations, wherein the carrier is aliquid, for administration, as for example, a nasal spray or as nasaldrops, include aqueous or oily solutions of the active ingredient.

Formulations suitable for nasal administration, wherein the carrier is asolid, include a coarse powder having a particle size, for example, inthe range of 20 to 500 microns which is administered in the manner inwhich snuff is administered (i.e., by rapid inhalation through the nasalpassage from a container of the powder held close up to the nose).Suitable powder compositions include, by way of illustration, powderedpreparations of the active ingredient thoroughly intermixed with lactoseor other inert powders acceptable for intrabronchial administration. Thepowder compositions can be administered via an aerosol dispenser orencased in a breakable capsule which may be inserted by the patient intoa device that punctures the capsule and blows the powder out in a steadystream suitable for inhalation.

Modulators may also be prepared in the form of suppositories (e.g., forrectal administration). Such compositions can be prepared by mixing thedrug with a suitable non-irritating excipient that is solid at ordinarytemperatures but liquid at the rectal temperature and will thereforemelt in the rectum to release the drug. Suitable excipients include, forexample, cocoa butter and polyethylene glycols.

Pharmaceutical compositions may be formulated as sustained releaseformulations (i.e., a formulation such as a capsule that effects a slowrelease of modulator following administration). Such formulations maygenerally be prepared using well known technology and administered by,for example, oral, rectal or subcutaneous implantation, or byimplantation at the desired target site. Carriers for use within suchformulations are biocompatible, and may also be biodegradable;preferably the formulation provides a relatively constant level ofmodulator release. The amount of modulator contained within a sustainedrelease formulation depends upon, for example, the site of implantation,the rate and expected duration of release and the nature of thecondition to be treated or prevented.

In addition to or together with the above modes of administration, amodulator may be conveniently added to food or drinking water (e.g., foradministration to non-human animals including companion animals (such asdogs and cats) and livestock). Animal feed and drinking watercompositions may be formulated so that the animal takes in anappropriate quantity of the composition along with its diet. It may alsobe convenient to present the composition as a premix for addition tofeed or drinking water.

Modulators are generally administered in a therapeutically effectiveamount. Preferred systemic doses range from about 0.1 mg to about 140 mgper kilogram of body weight per day (about 0.5 mg to about 7 g perpatient per day), with oral doses generally being about 5-20 fold higherthan intravenous doses. The amount of active ingredient that may becombined with the carrier materials to produce a single dosage form willvary depending upon the host treated and the particular mode ofadministration. Dosage unit forms will generally contain between fromabout 1 mg to about 500 mg of an active ingredient.

Packaged pharmaceutical compositions are also provided herein,comprising a C5a receptor modulatory amount of at least one C5a receptorantagonist in a container (preferably sealed) and instructions for usingthe C5a receptor antagonist to treat a condition responsive to C5areceptor modulation (e.g., rheumatoid arthritis, psoriasis,cardiovascular disease, reperfusion injury, bronchial asthma, chronicpulmonary obstructive disorder (COPD), cystic fibrosis, Alzheimer'sdisease, stroke, myocardial infarction, atherosclerosis, ischemic heartdisease or ischemia-reperfusion injury). The active agent(s) may beformulated for administration in a single pharmaceutical preparation(e.g., within the same pharmaceutical composition). Alternatively, eachof the active agents may be formulated for separate administration, bythe same or different routes of administration. Within a packagedpharmaceutical preparation, a C5a receptor modulatory amount may bepackaged as a single dose unit; alternatively, multiple doses may bepackaged together for convenience. The C5a receptor modulator may bepresented in any suitable container including, but not limited to, aplastic, paper, metal or glass package such as an ampoule, bottle, vial,blister package, infusion bag, syringe, inhaler or tube. For example, apackaged pharmaceutical preparation for oral administration of an activeagent may comprise a blister package containing rows of tablets.Instructions may be present on a label attached to the container or onexterior packaging, or may be provided as a package insert.

Methods of Use

C5a modulators provided herein may be used as agonists or (preferably)antagonists, such as inverse agonists, of C5a receptors in a variety ofcontexts, both in vitro and in vivo. Within certain aspects, C5aantagonists may be used to inhibit the binding of C5a receptor ligand(e.g., C5a) to C5a receptor in vitro or in vivo. In general, suchmethods comprise the step of contacting a C5a receptor with a sufficientconcentration of one or more C5a receptor modulators as provided herein,in the presence of C5a receptor ligand in aqueous solution and underconditions otherwise suitable for binding of the ligand to C5a receptor.The C5a receptor may be present in suspension (e.g., in an isolatedmembrane or cell preparation), or in a cultured or isolated cell. Withincertain embodiments, the C5a receptor is expressed by a cell present ina patient, and the aqueous solution is a body fluid. In general, theconcentration of C5a receptor modulator contacted with the receptorshould be sufficient to inhibit C5a binding to C5a receptor in vitro asmeasured, for example, using a calcium mobilization assay or chemotaxisassay as described herein.

Also provided herein are methods for modulating, preferably inhibiting,the signal-transducing activity of a C5a receptor. Such modulation maybe achieved by contacting a C5a receptor (either in vitro or in vivo)with a C5a receptor modulatory amount of one or more C5a receptormodulators provided herein under conditions suitable for binding of themodulator(s) to the receptor. The receptor may be present in solution orsuspension, in a cultured or isolated cell preparation or within apatient. Modulation of signal transducing activity may be assessed bydetecting an effect on calcium ion conductance (also referred to ascalcium mobilization or flux) or by detecting an effect on C5areceptor-mediated cellular chemotaxis. C5a receptor modulator(s)provided herein are preferably administered to a patient (e.g., a human)orally or topically, and are present within at least one body fluid ofthe animal while modulating C5a receptor signal-transducing activity.

The present invention further provides methods for treating patientssuffering from conditions responsive to C5a receptor modulation. As usedherein, the term “treatment” encompasses both disease-modifyingtreatment and symptomatic treatment, either of which may be prophylactic(i.e., before the onset of symptoms, in order to prevent, delay orreduce the severity of symptoms) or therapeutic (i.e., after the onsetof symptoms, in order to reduce the severity and/or duration ofsymptoms). A condition is “responsive to C5a receptor modulation” ifmodulation of C5a receptor activity results in alleviation of thecondition or a symptom thereof. Patients may include primates(especially humans), domesticated companion animals (such as dogs, cats,horses) and livestock (such as cattle, pigs, sheep), with dosages asdescribed herein.

Conditions that are responsive to C5a receptor modulation include thefollowing:

Autoimmune disorders—e.g., rheumatoid arthritis, systemic lupuserythematosus (and associated glomerulonephritis), psoriasis, Crohn'sdisease, vasculitis, irritable bowel syndrome, dermatomyositis, multiplesclerosis, bronchial asthma, pemphigus, pemphigoid, scleroderma,myasthenia gravis, autoimmune hemolytic and thrombocytopenic states,Goodpasture's syndrome (and associated glomerulonephritis and pulmonaryhemorrhage), immunovasculitis, tissue graft rejection, and hyperacuterejection of transplanted organs.

For asthma therapy, C5a receptor antagonists provided herein may be usedto prevent or decrease the severity of both acute early phase asthmaattack and the late phase reactions that follow such an asthma attack.

Inflammatory disorders and related conditions—e.g., neutropenia, sepsis,septic shock, Alzheimer's disease, stroke, inflammation associated withsevere burns, lung injury, and ischemia-reperfusion injury,osteoarthritis, as well as acute (adult) respiratory distress syndrome(ARDS), chronic pulmonary obstructive disorder (COPD), systemicinflammatory response syndrome (SIRS), cystic fibrosis, and multipleorgan dysfunction syndrome (MODS). Also included are pathologicsequellae associated with insulin-dependent diabetes mellitus (includingdiabetic retinopathy), lupus nephropathy, Heyman nephritis, membranousnephritis and other forms of glomerulonephritis, contact sensitivityresponses, and inflammation resulting from contact of blood withartificial surfaces that can cause complement activation, as occurs, forexample, during extracorporeal circulation of blood (e.g., duringhemodialysis or via a heart-lung machine, for example, in associationwith vascular surgery such as coronary artery bypass grafting or heartvalve replacement) such as extracorporeal post-dialysis syndrome, or inassociation with contact with other artificial vessel or containersurfaces (e.g., ventricular assist devices, artificial heart machines,transfusion tubing, blood storage bags, plasmapheresis,plateletpheresis, and the like).

Cardiovascular and Cerebrovascular Disorders—e.g., myocardialinfarction, coronary thrombosis, vascular occlusion, post-surgicalvascular reocclusion, atherosclerosis, traumatic central nervous systeminjury, and ischemic heart disease. For example, a C5a receptormodulatory amount of a compound provided herein may be administered to apatient at risk for myocardial infarction or thrombosis (i.e., a patientwho has one or more recognized risk factor for myocardial infarction orthrombosis, such as, but not limited to, obesity, smoking, high bloodpressure, hypercholesterolemia, previous or genetic history ofmyocardial infarction or thrombosis) in order reduce the risk ofmyocardial infarction or thrombosis.

HIV infection and AIDS—C5a receptor modulators provided herein may beused to inhibit HIV infection, delay AIDS progression or decrease theseverity of symptoms of HIV infection and AIDS.

In a further aspect, C5a receptor modulators may be used to perfuse adonor organ prior to transplantation of the organ into a recipientpatient. Such perfusion is preferably carried out using a solution(e.g., pharmaceutical composition) comprising a concentration of themodulator that is sufficient to inhibit C5a receptor-mediated effects invitro and/or in vivo. Such perfusion preferably reduces the severity orfrequency of one or more of the inflammatory sequelae following organtransplantation when compared to that occurring in control (including,without restriction, historical control) transplant recipients who havereceived transplants of donor organs that have not been so perfused.

Within further aspects, C5a antagonists provided herein may be used totreat Alzheimer's disease, multiple sclerosis, and cognitive functiondecline associated with cardiopulmonary bypass surgery and relatedprocedures. Such methods comprise administration of a therapeuticallyeffective amount of a C5a antagonist provided herein to a patientafflicted with one or more of the above conditions, or who is consideredto be at risk for the development of one or more such conditions.

Suitable patients include those patients suffering from or susceptibleto a disorder or disease identified herein. Typical patients fortreatment as described herein include mammals, particularly primates,especially humans. Other suitable patients include domesticatedcompanion animals such as a dog, cat, horse, and the like, or alivestock animal such as cattle, pig, sheep and the like.

In general, treatment methods provided herein comprise administering toa patient a C5a receptor modulatory amount of one or more compounds orforms thereof provided herein. Treatment regimens may vary depending onthe compound used and the particular condition to be treated; fortreatment of most disorders, a frequency of administration of 4 timesdaily or less is preferred. In general, a dosage regimen of 2 timesdaily is more preferred, with once a day dosing particularly preferred.It will be understood, however, that the specific dose level andtreatment regimen for any particular patient will depend upon a varietyof factors including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,route of administration, rate of excretion, drug combination (i.e.,other drugs being administered to the patient) and the severity of theparticular disease undergoing therapy, as well as the judgment of theprescribing medical practitioner. In general, the use of the minimumdose sufficient to provide effective therapy is preferred. Patients maygenerally be monitored for therapeutic effectiveness using medical orveterinary criteria suitable for the condition being treated orprevented.

As noted above, certain compounds and compositions provided herein areuseful as inhibitors of C5a receptor-mediated chemotaxis (e.g., they maybe used as standards in assays of such chemotaxis). Accordingly, methodsare provided herein for inhibiting C5a receptor-mediated cellularchemotaxis, preferably leukocyte (e.g., neutrophil) chemotaxis. Suchmethods comprise contacting white blood cells (particularly primatewhite blood cells, especially human white blood cells) with one or morecompounds provided herein. Preferably the concentration is sufficient toinhibit chemotaxis of white blood cells in an in vitro chemotaxis assay,so that the levels of chemotaxis observed in a control assay aresignificantly higher, as described above, than the levels observed in anassay to which a compound as described herein has been added.

Dosage levels of the order of from about 0.1 mg to about 140 mg perkilogram of body weight per day are useful in the treatment orprevention of conditions involving pathogenic C5a activity (about 0.5 mgto about 7 g per human patient per day). The amount of active ingredientthat may be combined with the carrier materials to produce a singledosage form will vary depending upon the host treated and the particularmode of administration. Dosage unit forms will generally contain betweenfrom about 1 mg to about 500 mg of an active ingredient. For compoundsadministered orally, transdermally, intravaneously, or subcutaneously,it is preferred that sufficient amount of the compound be administeredto achieve a serum concentration of 5 ng (nanograms)/mL-10 μg(micrograms)/mL serum, more preferably sufficient C5a receptor modulatorto achieve a serum concentration of 20 ng-1 μg/mL serum should beadministered, most preferably sufficient C5a receptor modulator toachieve a serum concentration of 50 ng/mL-200 ng/mL serum should beadministered. For direct injection into the synovium (for the treatmentof arthritis) sufficient C5a receptor modulator should be administeredto achieve a local concentration of approximately 1 micromolar.

Frequency of dosage may also vary depending on the compound used and theparticular disease treated. However, for treatment of most disorders, adosage regimen of 4 times daily, three times daily, or less ispreferred, with a dosage regimen of once daily or 2 times daily beingparticularly preferred. It will be understood, however, that thespecific dose level for any particular patient will depend upon avariety of factors including the activity of the specific compoundemployed, the age, body weight, general health, sex, diet, time ofadministration, route of administration, and rate of excretion, drugcombination (i.e., other drugs being administered to the patient), theseverity of the particular disease undergoing therapy, and otherfactors, including the judgment of the prescribing medical practitioner.

Within separate aspects, the present invention provides a variety ofnon-pharmaceutical in vitro and in vivo uses for the compounds providedherein. For example, such compounds may be labeled and used as probesfor the detection and localization of C5a receptor (in samples such ascell preparations or tissue sections, preparations or fractionsthereof). Compounds may also be used as positive controls in assays forC5a receptor activity, as standards for determining the ability of acandidate agent to bind to C5a receptor, or as radiotracers for positronemission tomography (PET) imaging or for single photon emissioncomputerized tomography (SPECT). Such methods can be used tocharacterize C5a receptors in living subjects. For example, a C5areceptor modulator may be labeled using any of a variety of well knowntechniques (e.g., radiolabeled with a radionuclide such as tritium, asdescribed herein), and incubated with a sample for a suitable incubationtime (e.g., determined by first assaying a time course of binding).Following incubation, unbound compound is removed (e.g., by washing),and bound compound detected using any method suitable for the labelemployed (e.g., autoradiography or scintillation counting forradiolabeled compounds; spectroscopic methods may be used to detectluminescent groups and fluorescent groups). As a control, a matchedsample containing labeled compound and a greater (e.g., 10-fold greater)amount of unlabeled compound may be processed in the same manner. Agreater amount of detectable label remaining in the test sample than inthe control indicates the presence of C5a receptor in the sample.Detection assays, including receptor autoradiography (receptor mapping)of C5a receptor in cultured cells or tissue samples may be performed asdescribed by Kuhar in sections 8.1.1 to 8.1.9 of Current Protocols inPharmacology (1998) John Wiley & Sons, New York.

Modulators provided herein may also be used within a variety of wellknown cell separation methods. For example, modulators may be linked tothe interior surface of a tissue culture plate or other support, for useas affinity ligands for immobilizing and thereby isolating, C5areceptors (e.g., isolating receptor-expressing cells) in vitro. Withinone preferred embodiment, a modulator linked to a fluorescent marker,such as fluorescein, is contacted with the cells, which are thenanalyzed (or isolated) by fluorescence activated cell sorting (FACS).

Preparation of Compounds

Representative methods for preparing compounds of Formula I and FormulaII are shown in Schemes 1-4. Those skilled in the art will recognizethat the reagents and synthetic transformations in the following Schemescan be readily modified to produce additional compounds of Formula I andFormula II. When a protecting group is required, an optionaldeprotection step may be employed. Suitable protecting groups andmethodology for protection and deprotection such as those described inProtecting Groups in Organic Synthesis by T. Greene are well known.Compounds and intermediates requiring protection/deprotection will bereadily apparent.

Abbreviations used in the following Schemes and Examples are as follows:

Ac₂O acetic anhydride BOPbenzotriazol-1-yloxy-tris(dimethylamino)-phosphonium hexafluorophospaten-BuLi n-butyl lithium CDCl₃ deuterated chloroform DCE 1,2-dichlorethaneDCM dichloromethane DEAD diethyl azidocarboxylate DIBAL-Hdiisobutylaluminum hydride DIEA diiosopropylethylamine DMAN,N-dimethylacetamide DMAP 4-N,N-dimethylaminopyridine DMFN,N-dimethylformamide DPPF 1,1′-bis(diphenylphosphino)ferrocene EtOAcethyl acetate HOAc acetic acid HPLC high pressure liquid chromatography¹H NMR proton nuclear magnetic resonance Hz hertz LAH lithium aluminumhydride LDA lithium diisopropylamide LC/MS liquid chromatography/massspectrometry MEK methyl ethyl ketone (2-butanone) MHz megahertz MS massspectrometry (M + 1) mass + 1 NMP N-methyl-2-pyrrolidone NBSN-bromosuccinimde δ chemical shift Pd(PPh₃)₄tetrakis(triphenylphosphine) palladium (0) POCl₃ phosphorous oxychloridePrMgCl n-propylmagnesium chloride PTLC preparative thin layerchromatography THF tetrahydrofuran TMSCN trimethylsilylcyanide 18-C-618-crown-6

Scheme 1 illustrates a method for preparing compounds of Formula I whereR₂ is NR₄R₅. Illustrative examples for this method are provided inExample 1 from WO 98/277066 which are hereby incorporated by reference.

Specific examples for the preparation of compounds of Formula I andFormula II (and the other Formulas provided herein) by the methodsillustrated in the above Schemes are provided in the following Examples.Unless otherwise specified all starting materials and reagents are ofstandard commercial grade, and are used without further purification, orare readily prepared from such materials by routine methods. Thoseskilled in the art of organic synthesis will recognize that startingmaterials and reaction conditions may be varied to achieve the desiredend product.

EXAMPLES Example 1 Preparation of Certain Starting Materials A.Synthesis of 2,6-diethylphenylboronic Acid

2,6-Diethyl bromobenzene (38.2 g, 180.2 mmol) is added dropwise throughan additional funnel over a 1 hour period to a solution of n-BuLi (2.0 Min cyclohexane, 99.1 mL, 198.2 mmol) in THF (380 mL) at −75° C. Afteraddition, the reaction mixture is stirred at −75° C. for 30 minutes;trimethyl borate (28.1 g, 270.3 mmol) is added slowly over a 40 minuteperiod. The reaction mixture is warmed to room temperature overnight. 2NHCl (250 mL) is added slowly and the resulting mixture is stirred for 1hour. The organic layer is separated and the aqueous layer is extractedwith ether (2×200 mL). The combined organic layers are dried overanhydrous Na₂SO₄ and the solvents are removed in vacuo. Hexane (400 mL)is added to the residue and a white precipitate is formed. Filtrationand drying in vacuo give 2,6-diethylphenyl boronic acid as a whitesolid. ¹H NMR: (CDCl₃) 7.22 (t, 1H), 7.04 (s, 2H), 4.65 (s, 2H), 2.64(q, 4H), 1.22 (t, 6H).

B. Synthesis of 2,6-dimethyl-3-methoxybenzeneboronic Acid

Step 1. Preparation of Aldehyde

A solution of 2-bromo-m-xylene (4.2 g, 23 mmol) in dichloromethane (5mL) at −78° C. is added dropwise to a solution of titanium tetrachloride(5.0 mL, 45 mmol) and dichloromethyl methyl ether (2.3 mL, 25 mmol) indichloromethane (20 mL). After the addition is complete, the mixture isallowed to warm to room temperature and stirred for and additional 4hours before being poured onto ice water. The reaction is extracted withdichloromethane. The organic fraction is washed with water, dried(Na₂SO₄), and concentrated to give the aldehyde as a pale yellow solid(4.7 g), which is used in the next step without further purification: ¹HNMR (CDCl₃) 10.1 (s, 1H), 7.68 (d, 1H), 7.22 (d, 1H), 2.79 (s, 3H), 2.45(s, 3H).

Step 2. Preparation of methyl ether

M-chloroperoxybenzoic acid (68%, 8.4 g, 33 mmol) is added to a solutionof the above aldehyde (4.7 g) in dichloromethane (120 mL). The mixtureis stirred at reflux overnight and concentrated in vacuo. The residue isdissolved in ethyl acetate and washed successively with saturated NaHCO₃(3 times), saturated NaHSO₃, and water. The organic fraction is dried(Na₂SO₄) and concentrated to give the crude formate (4.4 g). The formateis treated with potassium carbonate (4 g) in ethanol (80 mL) at roomtemperature for 20 minutes, followed by filtration and concentration togive the corresponding alcohol. The crude alcohol is dissolved inacetone (160 mL) and dimethyl sulfate (2.7 mL, 29 mmol), and potassiumcarbonate (8.0 g, 58 mmol) is added. The mixture is stirred at refluxfor 5 hours. After cooling to room temperature, filtration,concentration, and flash chromatography provide the desired methyl etheras a colorless oil (3.3 g). ¹H NMR (CDCl₃) 7.02 (d, 1H), 6.73 (d, 1H),3.80 (s, 3H), 2.37 (s, 3H), 2.35 (s, 3H).

Step 3. Preparation of 2,6-dimethyl-3-methoxybenzeneboronic Acid

A solution of 2,4-dimethyl-3-bromoanisole (3.3 g, 15 mmol) in THF (15mL) is added dropwise at −78° C. to a solution of n-butyllithium (11 mLof 1.6M in hexane, 17 mmol) in THF (35 mL). After 30 minutes, trimethylborate (2.3 mL, 20 mmol) is added and the mixture is allowed to warm toroom temperature overnight. The mixture is poured onto 10% HCl andextracted with ethyl acetate. The organic fraction is washed withsaturated brine, dried (Na₂SO₄), and concentrated to give the desiredproduct as a brownish oil. ¹H NMR (CDCl₃) 6.98 (d, 1H), 6.75 (d, 1H),4.64 (br s), 3.80 (s, 3H), 2.2.7 (s, 3H), 2.22 (s, 3H).

F. Synthesis of (S)-methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine

Ethyl chloroformate (7.74 g, 71.3 mmol) is added dropwise to a mixtureof (S)-1,2,3,4-tetrahydro-naphthalen-1-ylamine (10.0 g, 67.9 mmol) andK₂CO₃ (18.8 g, 136 mmol) in CH₃CN (100 mL). The resulting mixture isstirred at room temperature overnight. Water (100 mL) is added and themixture is extracted with ether (2×100 mL). The combined extract iswashed with 1 N HCl (2×10 mL), water, dried (Na₂SO₄), and concentratedin vacuo to give (S)-(1,2,3,4-tetrahydro-naphthalen-1-yl)-carbamic acidethyl ester as a solid.

(1,2,3,4-Tetrahydro-naphthalen-1-yl)-carbamic acid ethyl ester (5.0 g,22.8 mmol) is added slowly under nitrogen to a suspension of LiAlH₄ (2.6g, 68 mmol) in THF (50 mL). The resulting mixture is heated at 75° C.with stirring for 2 hours. On cooling, Na₂SO₄.10H₂O (15.0 g) and ether(100 mL) are added to the mixture. The resulting mixture is stirred atroom temperature for 1 hour, filtered through celite, and concentratedin vacuo. 1 N HCl (20 mL) and ether (20 ML) are added to the residue.The organic layer is separated and discarded. The aqueous layer isbasified with 1 N NaOH and extracted with CH₂Cl₂ (2×25 mL). The combinedextract is washed with water (2×), dried (Na₂SO₄) and concentrated togive (S)-methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine as an oil.[α]^(RT)=−10.6 (0.02, EtOH). ¹H NMR (CDCl₃) 7.30 (m, 1H), 7.06-7.20 (m,3H), 3.66 (t, 1H), 2.78 (m, 2H), 2.50 (s, 3H), 1.70-2.00 (m, 4H).

Similar procedures are applied in the synthesis of the following amines:

-   (R)-Methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine;-   (S)-Ethyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine;-   (S)-Propyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine;-   (S)-Indan-1-yl-methyl-amine;-   (±)Methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine; and-   (±)Indan-1-yl-methyl-amine.

G. Synthesis of 5-methylindole-4-boronic Acid

Fuming nitric acid (>90% yellow fuming HNO₃) is slowly added to asolution of 2-bromo-m-xylene (20 g, 150 mmol) in acetic acid (100 mL)cooled in an ice bath (above freezing point). The resulting mixture isallowed to warm to room temperature, stirred for 1 hour, and heated at80° C. for 2 hours or until the reaction is complete by GC/MS analysisfollowing micro-scale base work-up. The reaction mixture is cooled toroom temperature and poured into ice/water with stirring. The resultingyellow precipitates are collected by suction filtration and air dried toobtain 2,6-dimethyl-3-nitrobromobenzene.

Bredereck's reagent (tert-butoxybis(dimethylamino)methane (16 g, 91mmol) is added to a solution of 2,6-dimethyl-3-nitrobromobenzene (20 g,87 mmol) in anhydrous DMF (120 mL) at room temperature. The reactionmixture is heated at 120-125° C. under N₂ for 5 hours or until startingmaterial is mostly consumed according to TLC. The reaction mixture isallowed to cool to room temperature, poured into water (300 mL), andextracted with dichloromethane (100 mL×3). The combined extracts aredried over anhydrous sodium sulfate, filtered, and concentrated toobtain a mixture of enamines as a dark brown oil. This material is usedin the next step without purification.

The crude mixture is dissolved in acetic acid/water (250 mL of 4:1),cooled to 0° C. and treated with zinc dust (57 g, 870 mmol) added slowlyin portions. After complete addition, the reaction mixture is heated at110° C. for 4 hours. Zinc is removed by filtration through a celite padand the filtrate is extracted with dichloromethane (100 mL×3). Thecombined extracts are dried over anhydrous sodium sulfate, concentrated,and purified by flash chromatography on silica gel (EtOAc/Hexane 1:20)to obtain 4-bromo-5-methylindole as a light purple oil.

A solution of 4-bromo-5-methylindole (800 mg, 3.8 mmol) in anhydrousether (8 mL) is added with stirring to a suspension of potassium hydride(560 mg, 4.2 mmol, 30% dispersion in mineral oil) in anhydrous ether at0° C. under argon. The resulting mixture is cooled to −78° C. andtert-butyllithium (4.9 mL of 1.7 M in pentane, 8.4 mmol) is slowlyadded. The resulting cream-colored mixture is stirred at −78° C. for 1hour. Tributylborate (3.1 mL, 11.4 mmol) is slowly added and thereaction mixture is stirred for 1 hour at −78° C. before being allowedto slowly warm to room temperature. More anhydrous ether is added tofacilitate stirring. After stirring for 24 hours, the resulting stickymixture is diluted with ether and transferred in portions with stirringto a precooled solution of 1 M phosphoric acid (50 mL). After stirringfor 30 minutes, the acidic mixture is extracted with diethyl ether (75mL×3) and the combined extracts are extracted with 1 N sodium hydroxide(20 mL×4). The combined base extracts are cooled with an ice bath,acidified with 1 M phosphoric acid and extracted with ethyl acetate (20mL×3). The combined extracts are washed with brine (20 mL), dried overanhydrous sodium sulfate, filtered and concentrated to obtain a beigeresidue. The residue is triturated with hexane to obtain the desired5-methylindole-4-boronic acid as a beige gum (230 mg).

H. Synthesis of 6-isopropyl-2-methyl-3-nitrobenzeneboronic Acid

6-Isopropyl-2-methylbenzeneboronic acid (8 g) is added portionwise over1 hour to 90% HNO₃ (50 mL) at −40° C., maintaining an internaltemperature below −30° C. After addition, the mixture is stirred at −40to −30° C. for 15 minutes, then poured onto ice, and diluted with water.The solid is collected by filtration, washed with water and dried togive 6-isopropyl-2-methyl-3-nitrobenzeneboronic acid as a white solid.NMR (DMSO-d6) 7.78 (d, 2H), 7.30 (d, 2H), 2.85 (m, 1H), 2.38 (s, 3H),1.15 (d, 6H).

I. Synthesis of 5-isopropyl-1H-indazole-4-boronic Acid

Step 1. Preparation of 4-Bromo-5-isopropyl-1H-indazole

Nitric acid (30 mL, fuming) is added slowly to an ice-cold solution of2-isopropyl-6-methyl-bromobenzene (10 g, 213 mmol) in acetic acid (60mL). The mixture is heated 1 hour at 90° C. and cooled to roomtemperature. The reaction mixture is poured into 200 mL ice-water andextracted with CH₂Cl₂ (3×60 mL). The combined extracts are washed with 1N NaOH (3×40 mL) and then water (40 mL), dried (Na₂SO₄), andconcentrated to yield crude 2-isopropyl-6-methyl-5-nitro-bromobenzenewhich is dissolved in AcOH (75 mL)/EtOH (75 mL). To this is added Fepower (5.3 g, 95 mmol) and the mixture is refluxed for 2 hours. Themixture is cooled to room temperature, diluted with water, andneutralized with solid Na₂CO₃. The mixture is extracted with EtOAc,dried (Na₂SO₄), and concentrated in vacuo. The residue is purified byflash chromatography (elution with Hex/EtOAc 4:1) to yield3-bromo-4-isopropyl-2-methyl-aniline. A solution of NaNO₂ (798 mg, 12mmol) in H₂O (10 mL) is added dropwise at 0° C. to a slurry of3-bromo-4-isopropyl-2-methyl-aniline (2.4 g, 11 mmol) in HBF₄ (15mL)-H₂O (15 mL), and the mixture is stirred for 1 hour at 0° C. Theresulting solid is filtered, washed with cold water and then Et₂O, anddried under reduced pressure to yield the diazonium salt as a beigesolid. The diazonium salt is added in one portion to mixture of KOAc(1.5 g, 15 mmol) and 18-C-6 (98 mg, 0.37 mmol) in ethanol-free CHCl₃ (70mL) at room temperature. The mixture is stirred for 1 hour and theresulting solid is removed by filtration. The filtrate is washed withwater, dried (Na₂SO₄), and concentrated in vacuo. The residue ispurified by flash chromatography (elution with Hex/EtOAc 4:1) to yield4-bromo-5-isopropyl-1H-indazole. NMR (CDCl₃) 8.03 (br s, 1H), 7.41 (d,1H), 7.35 (d, 1H), 3.55 (m, 1H), 1.24 (d, 6H).

Step 2. Preparation of 5-Isopropyl-1H-indazole-4-boronic Acid

A solution of 4-bromo-5-isopropyl-1H-indazole (1.6 g, 6.9 mmol) in Et₂O(4 mL) is added slowly to a suspension of KH (1.0 g of 30% dispersion inmineral oil, 7.7 mmol) in Et₂O (20 mL) at 0° C. and the mixture isstirred for 20 minutes. After cooling to −78° C., t-BuLi (8.9 mL of 1.7M in Hex, 15 mmol) is added and the resulting mixture is stirred for 40minutes at −78° C. To this is added B(On-Bu)₃ (5.6 mL, 21 mmol) and themixture is stirred for 24 hours at room temperature. The reactionmixture is quenched with 1N H₃PO₄ and extracted with Et₂O. The combinedEt₂O layers are back-extracted with 1N NaOH (3×10 mL). The combined NaOHextracts are acidified with 1N H₃PO₄ and extracted with EtOAc. The EtOAcextracts are washed with saturated brine, dried (MgSO₄), andconcentrated to yield 5-isopropyl-1H-indazole-4-boronic acid. ¹H NMR(CDCl₃) 7.85 (s, 1H), 7.42 (d, 1H), 7.37 (d, 1H), 3.6 (br s, 2H), 2.88(m, 1H), 1.32 (d, 6H).

J. Synthesis of 3-isopropyl-1H-indazole-4-boronic Acid

Step 1. Preparation of 1-(2-Bromo-6-fluoro-phenyl)-2-methyl-propan-1-one

To a solution of n-BuLi (25 mL of 1.6 M solution in hexane, 40 mmol) inTHF (100 mL) is added 2,2,6,6-teramethylpiperidine (6.8 mL, 40 mmol) at−78° C. and the mixture is stirred for 20 minutes. To this is added3-bromofluorobenzene (7.0 g, 40 mmol). After stirring for 3 hours at−78° C., DMF (15 mL, 200 mmol) is added and the mixture is warmed toroom temperature and stirred for 1 hour. The mixture is quenched with 1NHCl and extracted with EtOAc. The combined extracts are dried (MgSO₄)and concentrated in vacuo. The residue is purified by flashchromatography (elution with Hex/EtOAc 10:1) to yield2-bromo-6-fluoro-benzaldehyde. ¹H NMR (CDCl₃) 10.4 (s, 1H), 7.48-7.39(m, 2H), 7.18-7.14 (m, 1H).

Isopropylmagnesium chloride (18 mL of 2 M in Et₂O, 35 mmol) is added toa solution of 2-bromo-6-fluoro-benzaldehyde (6.0 g, 30 mmol) in THF (40mL) at −78° C. and the mixture is stirred for 1 hour at 0° C. Themixture is poured into saturated NH₄Cl and extracted with EtOAc. Theresulting crude alcohol is oxidized directly by Swern oxidation to yield1-(2-bromo-6-fluoro-phenyl)-2-methyl-propan-1-one. ¹H NMR (CDCl₃) 7.38(d, 1H), 7.22 (m, 1H), 7.03 (t, 1H), 3.10 (m, 1H), 1.11 (d, 6H).

Step 2. Preparation of 3-Isopropyl-1H-indazole-4-boronic acid

A mixture of 1-(2-bromo-6-fluoro-phenyl)-2-methyl-propan-1-one (1.1 g,4.5 mmol) and anhydrous hydrazine (0.17 mL, 5.4 mmol) in ethylene glycol(10 mL) is heated for 16 hours at 160° C. Water is added and the mixtureis extracted with CH₂Cl₂. The combined extracts are dried (MgSO₄) andconcentrated in vacuo. The residue is purified by flash chromatographyto yield 4-bromo-3-isopropyl-1H-indazole. ¹H NMR (CDCl₃) 10.1 (br s,1H), 7.38 (d, 1H), 7.32 (d, 1H), 7.17 (t, 1H), 3.99 (m, 1H), 1.43 (d,6H).

4-Bromo-3-isopropyl-1H-indazole is converted to the correspondingboronic acid following analogous procedures to that given in thepreceding example. ¹H NMR (CD₃OD) 7.44 (d, 1H), 7.32 (t, 1H), 7.05 (d,1H), 3.56 (m, 1H), 1.38 (d, 6H). LCMS (m/z): 205.45 (MH)⁺.

Example 2 Synthesis of1-(5-isopropyl-1H-indazol-4-yl)-4-(5-isopropyl-2-methyl-phenoxymethyl)-3-methyl-isoquinoline

Step 1. Preparation of 1-Chloro-3-methyl-isoquinoline-4-carbaldehyde

n-BiLi (12.3 ml of 1.6 M in hexane, 20 mmol) is added dropwise to asolution of 4-bromo-1-chloro-3-methyl-isoquinoline (4.6 g, 18 mmol) inTHF (80 ml) at −78° C. and the mixture is stirred for 40 min. DMF (4.2ml, 54 mmol) is then added slowly. The cold bath is removed, andstirring is continued for 15 min. The mixture is acidified with 1N—HClto pH 2 and extracted with ether. The combined extracts are dried overNa₂SO₄ and concentrated in vacuo. The residue is chromatographed onsilica gel to give 3.0 g of1-chloro-3-methyl-isoquinoline-4-carbaldehyde. ¹H NMR (CDCl₃) 10.9 (s,1H), 9.06 (d, 1H), 8.40 (d, 1H), 7.88 (t, 1H), 7.70 (t, 1H), 2.99 (s,3H).

Step 2. Preparation of 1-Chloro-4-chloromethyl-3-methyl-isoquinoline

LAH (3.6 ml of 1M in THF, 3.62 mmol) is added to a solution of1-chloro-3-methyl-isoquinoline-4-carbaldehyde (744 mg, 3.62 mmol) in THF(10 ml) at 0° C. After stirring for 30 min at ambient temperature, themixture is quenched with small amounts of saturated sodium sulfate andfiltered through a pad of celite. The filtrate is concentrated in vacuoto give the crude alcohol. SOCl₂ (13 ml of 2M in DCM, 25 mmol) is addedto a solution of the crude alcohol in DCM (5 ml). After stirring for 1 hat room temperature, the mixture is concentrated in vacuo and theresidue is chromatographed on silica gel to give 737 mg of1-chloro-4-chloromethyl-3-methyl-isoquinoline. ¹H NMR (CDCl₃) 8.44 (d,1H), 8.18 (d, 1H), 8.02 (t, 1H), 7.82 (t, 1H), 5.03 (s, 2H), 3.00 (s,3H).

Step 3. Preparation of1-(5-Isopropyl-1H-indazol-4-yl)-4-(5-isopropyl-2-methyl-phenoxymethyl)-3-methyl-isoquinoline

A solution of 1-chloro-4-chloromethyl-3-methyl-isoquinoline (150 mg,0.66 mmol) is added to a mixture of carvacrol (199 mg, 1.33 mmol) andCsCO₃ (645 mg, 1.98 mmol) in DMF (8 ml). After stirring overnight atroom temperature, water is added and the mixture is extracted withether. The combined extracts are washed with saturated brine, dried overNa₂SO₄, and concentrated in vacuo. The residue is chromatographed onsilica gel to give 220 mg of1-chloro-4-(5-isopropyl-2-methyl-phenoxymethyl)-3-methyl-isoquinoline. Amixture of1-chloro-4-(5-isopropyl-2-methyl-phenoxymethyl)-3-methyl-isoquinoline(220 mg, 0.65 mmol), 5-isopropylindazole-4-boronic acid (173 mg, 0.85mmol), Pd(PPh₃)₄ (38 mg, 0.033 mmol), and Na₂CO₃ (1.3 ml of 2M in H₂O)in dioxane (8 ml) The mixture is heated for 18 h at 100° C. Aftercooling to room temperature, water is added and the resulting mixture isextracted with EtOAc. The combined extracts are dried over Na₂SO₄ andconcentrated in vacuo. The residue is chromatographed on silica gel togive 133 mg of1-(5-isopropyl-1H-indazol-4-yl)-4-(5-isopropyl-2-methyl-phenoxymethyl)-3-methyl-isoquinoline

¹H NMR (CDCl₃) 10.6 (br s, 1H), 8.18 (d, 1H), 7.68 (t, 1H), 7.54 (d,1H), 7.53 (s, 2H), 7.42 (s, 1H), 7.36 (t, 1H), 7.14 (d, 1H), 7.06 (s,1H), 6.86 (d, 1H), 5.59 (s, 2H), 3.01-2.94 (m, 1H), 2.91 (s, 3H),2.78-2.71 (m, 1H), 2.20 (s, 3H), 1.32 (d, 6H), 1.22 (d, 3H), 1.13 (d,3H).

LCMS (m/z):464.3 (MH)⁺

Example 3 Synthesis ofS-1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinolin-4-ylmethyl]-methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine

A mixture of4-chloromethyl-1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinoline (69mg, 0.2 mmol), S-methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine (97mg, 0.6 mmol), and K₂CO₃ (138 mg, 1 mmol) in MeCN (5 ml) is heated for16 h at 80° C. After cooling, water is added and the resulting mixtureis extracted with EtOAc. The combined extracts are dried over NaSO₄ andevaporated. The residue is chromatographed on silica gel eluting withEtOAc/hexane (1:1) to give 48 mg of[1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinolin-4-ylmethyl]-methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amine.

¹H NMR (CDCl3) 8.41 (d, 1H), 7.73-7.61 (m, 2H), 7.53-7.47 (m, 3H),7.38-7.31 (m, 2H), 7.13-7.07 (m, 3H), 4.30 (AB q, 2H), 4.18-4.13 (m,1H), 2.94-2.68 (m, 3H), 2.91 (s, 3H), 2.19 (s, 3H), 2.16-1.84 (m, 3H),1.20 (d, 3H), 1.11 (d, 3H).

LCMS (m/z):475.4 (MH)⁺

Example 4 Synthesis of 4-(2,2-dimethyl-morpholin-4-ylmethyl)-1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinolin-6-ol

Step 1. Preparation of 6-Methoxy-3-methyl-2H-isoquinolin-1-one

A mixture of 4-methoxy-2-methylbenzoic acid (10 g, 60 mmol) and dimethylcarbonate (10 ml, 120 mmol) in THF (80 ml) is added dropwise to LDA (120ml of 2M in heptane/THF/ethylbenzene) at −78° C. The cold bath isremoved, and after 4 h, water (100 ml) is added, and the resultingmixture is stirred overnight. After removal of organic solvents, theresidue is acidified with c-HCl to pH 2. The resulting solid isfiltered, dissolved in 1N—NaOH (80 ml), and then washed with ether (2×30ml). The aqueous layer is acidified with c-HCl and the resulting whitesolid is filtered to give 12 g of 2-carboxymethyl-4-methoxy-benzoicacid. ¹H NMR (DMSO) 12.3 (br s, 1H), 7.88 (d, 1H), 6.91 (d, 1H), 6.88(s, 1H), 3.90 (s, 2H), 3.79 (s, 3H).

Pyridine (5.0 ml, 62 mmol) is added slowly to suspension of2-carboxymethyl-4-methoxy-benzoic acid (10 g, 48 mmol) in aceticanhydride (80 ml) at 0° C. After stirring for 16 h, ether (100 ml) isadded. The resulting solid is collected and dried to give 10 g of4-acetyl-6-methoxy-isochroman-1,3-dione.

A suspension of 4-acetyl-6-methoxy-isochroman-1,3-dione (10 g, 43 mmol)in NH₄OH (60 ml) in a pressure vessel is heated for 2 h at 100° C. Aftercooling, the reaction mixture is poured into water. The white solid isfiltered, washed with water, and dried to give 6.7 g of6-methoxy-3-methyl-2H-isoquinolin-1-one. ¹H NMR (CDCl₃) 11.0 (br s, 1H),8.28 (d, 1H), 6.98 (d, 1H), 6.81 (s, 1H), 6.23 (s, 1H), 3.89 (s, 3H),2.37 (s, 3H).

Step 2. Preparation of1-(5-Isopropyl-1H-indazol-4-yl)-6-methoxy-3-methyl-isoquinoline-4-carbaldehyde

NBS (6.9 g, 40 mmol) is added in one portion to ice-cold suspension of6-methoxy-3-methyl-2H-isoquinolin-1-one (6.7 g, 35 mmol) in AcOH (100ml). The ice bath is removed, and after 1 h, the mixture is concentratedin vacuo. The residue is diluted with water (40 ml) and basified with10N—NaOH to pH 12. The resulting white solid is filtered and dried togive 9.3 g of 4-bromo-6-methoxy-3-methyl-2H-isoquinolin-1-one. ¹H NMR(CDCl₃) 8.10 (d, 1H), 7.14 (s, 1H), 7.08 (d, 1H), 3.89 (s, 3H), 2.37 (s,3H).

A suspension of 4-bromo-6-methoxy-3-methyl-2H-isoquinolin-1-one (9.3 g,35 mmol) in POCl₃ (50 ml) is refluxed for 1 h. After cooling to roomtemperature, POCl₃ is removed. The residue is diluted with water (50 ml)and neutralized with saturated NaHCO₃. The resulting solid is filteredand dried to give 9.8 g of4-bromo-1-chloro-6-methoxy-3-methyl-isoquinoline. ¹H NMR (CDCl₃) 8.17(d, 1H), 7.43 (s, 1H), 7.25 (d, 1H), 4.01 (s, 3H), 2.80 (s, 3H).

n-BuLi (12 ml of 1.6 M in hexane) is added dropwise to a solution of4-bromo-1-chloro-6-methoxy-3-methyl-isoquinoline (4.8 g, 17 mmol) in THF(60 ml) at −78° C. After stirring for 40 min, DMF (4 ml, 51 mmol) isadded slowly. The reaction is allowed to warm to 0° C. and the stirringis continued for 1 h. The mixture is then quenched with 1N—HCl (30 ml)and extracted with EtOAc. The combined extracts are dried over Na₂SO₄and evaporated. The residue is chromatographed on silica gel elutingwith EtOAc/hexane (1:4) to give 2.9 g of1-chloro-6-methoxy-3-methyl-isoquinoline-4-carbaldehyde. ¹H NMR (CDCl₃)10.8 (s, 1H), 8.61 (s, 1H), 8.24 (d, 1H), 7.29 (d, 1H), 4.01 (s, 3H),2.99 (s, 3H).

A mixture of 1-chloro-6-methoxy-3-methyl-isoquinoline-4-carbaldehyde(670 mg, 2.84 mmol), 5-isopropylindazole-4-boronic acid (580 mg, 2.84mmol), Pd(PPh₃)₄ (164 mg, 0.14 mmol), and Na₂CO₃ (4.3 ml of 2M in H₂O)in dioxane (15 ml) is heated for 18 h at 100° C. Usual work-up followedby flash chromatography over silica gel gives 342 mg of1-(5-isopropyl-1H-indazol-4-yl)-6-methoxy-3-methyl-isoquinoline-4-carbaldehyde.¹H NMR (CDCl₃) 10.9 (s, 1H), 8.68 (s, 1H), 7.65-7.37 (m, 3H), 7.42 (d,1H), 7.01 (d, 1H), 4.01 (s, 3H), 3.15 (s, 3H), 2.71-2.63 (m, 1H), 1.22(d, 3H), 1.10 (d, 3H).

Step 3. Preparation of4-(2,2-Dimethyl-morpholin-4-ylmethyl)-1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinolin-6-ol

BBr₃ (8.8 g, 35 mmol) is added slowly to a solution of1-(5-Isopropyl-1H-indazol-4-yl)-6-methoxy-3-methyl-isoquinoline-4-carbaldehyde(842 mg, 2.3 mmol) in DCM (15 ml) at −78° C. The cold bath is removedand the mixture is warmed to room temperature, and then stirred for afurther 1 h. The reaction mixture is poured into ice-water mixture andtreated with NH₄OH (6 ml). The layers are separated and the aqueouslayer is extracted with DCM. The combined organic layers are dried(Na₂SO₄) and evaporated. The residue is chromatographed on silica geleluting with EtOAc/MeOH (20:1) to give 490 mg of starting material and276 mg of6-hydroxy-1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinoline-4-carbaldehyde.¹H NMR (CDCl₃) 10.9 (s, 1H), 8.87 (s, 1H), 7.62-7.42 (m, 4H), 6.99 (d,1H), 3.09 (s, 3H), 2.73-2.67 (m, 1H), 1.23 (d, 3H), 1.12 (d, 3H).

A mixture of6-hydroxy-1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinoline-4-carbaldehyde(246 mg, 0.71 mmol), 2,2-dimethyl-morpholine hydrochloride (216 mg, 1.4mmol), NaB(OAc)₃H (451 mg, 2.1 mmol), and AcOH (30 uL) in DCM (10 ml) isstirred for 6 h at room temperature. 1N—NaOH (5 ml) is added and themixture is extracted with DCM. The combined extracts are dried overNa₂SO₄ and concentrated in vacuo. The residue is chromatographed onsilica gel eluting with DCM/MeOH (10:1) to give 207 mg of4-(2,2-dimethyl-morpholin-4-ylmethyl)-1-(5-isopropyl-1H-indazol-4-yl)-3-methyl-isoquinolin-6-ol.

¹H NMR (CD₃OD) 7.69-7.56 (m, 3H), 7.28-7.27 (m, 2H), 6.93 (d, 1H), 3.89(s, 2H), 3.72 (t, 2H), 2.72 (s, 3H), 2.63-2.43 (m, 5H), 1.23 (s, 6H),1.18 (d, 3H), 1.10 (d, 3H).

LCMS (m/z):445.5 (MH)⁺

Example 5 Synthesis of3-[4-[1-(benzyl-ethyl-amino)-butyl]-isoquinolin-1-yl}-2,4-dimethyl-phenol

Step 1. Preparation of1-(3-Methoxy-2,6-dimethyl-phenyl)-isoquinoline-4-carbaldehyde

Allyl chloroformate (2.7 ml, 25 mmol) is added dropwise at 0° C. to acooled mixture of isoquinoline (2.9 g, 23 mmol) and2,6-dimethyl-3-methoxyphenylmagnesium bromide (prepared from 7.3 g of2,6-dimethyl-3-methoxyphenyl bromide and 2.8 g of magnesium turnings) inTHF (30 ml). After stirring for 2 h at room temperature, the reactionmixture is quenched with saturated NH₄Cl and extracted with Et₂O. Thecombined extracts are washed with saturated brine, dried over Na₂SO₄,and concentrated to give 6.9 g of1-(3-methoxy-2,6-dimethyl-phenyl)-1H-isoquinoline-2-carboxylic acidallyl ester.

POCl₃ (9.4 ml, 100 mmol) is added slowly to DMF (15 ml, 200 mmol) in DCM(20 ml) at 0° C. and the mixture is stirred for 20 min. To this is addeda solution of the above prepared compound (6.9 g, 20 mmol) in DCM (10ml) at 0° C. After stirring overnight at room temperature, the resultingmixture is quenched with saturated NH₄Cl and then extracted with DCM.The combined extracts are washed with saturated brine, dried overNa₂SO₄, and concentrated to give4-formyl-1-(3-methoxy-2,6-dimethyl-phenyl)-1H-isoquinoline-2-carboxylicacid allyl ester which is dissolved in DCM (50 ml). To this are addedPd(PPh₃)₄ (60 mg, 0.052 mmol) and morpholine (2 ml, 23 mmol). Afterstirring for 1 h at room temperature, the mixture is diluted with DCM(50 ml) and cool to 0° C. DDQ (4.9 g, 22 mmol) is added portionwise andthe mixture is stirred for 30 min at 0° C. The reaction mixture isslowly poured into a solution of saturated NaHCO₃ and extracted withDCM. The combined extracts are washed with saturated brine and driedover Na₂SO₄. The organics are concentrated, and the residue ischromatographed on silica gel to give 4.9 g of1-(3-methoxy-2,6-dimethyl-phenyl)-isoquinoline-4-carbaldehyde.

¹H NMR (CD₃Cl) 10.4 (s, 1H), 9.30 (d, 1H), 9.05 (s, 1H), 7.89 (t, 1H),7.67 (d, 1H), 7.58 (t, 1H), 7.15 (d, 1H), 6.92 (d, 1H), 3.88 (s, 3H),1.82 (s, 3H), 1.76 (s, 3H).

Step 2. Preparation ofBenzyl-ethyl-{1-[1-(3-methoxy-2,6-dimethyl-phenyl)-isoquinolin-4-yl]-butyl}-amine

A mixture of the aldehyde from Step 1 (115 mg, 0.39 mmol), benzotriazole(50 mg, 0.41 mmol), and EtOH (0.5 ml) in toluene (8 ml) is heated toreflux for 2 h under a Dean-Stark water trap. The solvents are removedcompletely by using IR-Dancer and the residue is dissolved in THF (6ml). After cooling to 0° C., n-PrMgCl (0.3 ml of 2M solution in Et₂O,0.59 mmol) is added slowly and the mixture is stirred for 15 min atambient temperature. Water is added and the mixture is extracted withEtOAc. The combined extracts are dried over Na₂SO₄ and concentrated. Theresidue is chromatographed on silica gel to give 130 mg ofbenzyl-ethyl-{1-[1-(3-methoxy-2,6-dimethyl-phenyl)-isoquinolin-4-yl]-butyl}-amineas a colorless oil.

¹H NMR (CD₃Cl) 1:1 mixture of atropisomers 8.63 (s, 1H), 8.43 (d, 1H),7.64 (t, 1H), 7.52 (d, 1H), 7.42 (t, 1H), 7.26-7.19 (m, 5H), 7.10 (d,1H), 6.88 (d, 1H), 4.47-4.42 (m, 1H), 3.87 (s, 3H), 3.68 (s, 2H),2.85-2.78 (m, 1H), 2.70-2.61 (m, 1H), 2.14-2.05 (m, 2H), 1.80 (2s, 3H),1.74 (2s, 3H), 1.28-1.19 (m, 2H), 1.02 (t, 3H), 0.92 (t, 3H).

Step 3. Preparation of3-{4-[1-Benzyl-ethyl-amino)-butyl]isoquinolin-1-yl}-2,4-dimethyl-phenol

BBr₃ (2 ml of 1M solution in DCM, 2.0 mmol) is added to a solution ofbenzyl-ethyl-{1-[1-(3-methoxy-2,6-dimethyl-phenyl)-isoquinolin-4-yl]-butyl}-amine(95 mg, 0.21 mmol) in DCM (8 ml) at −78° C. The mixture is allowed towarm to room temperature slowly, and after 2h, the mixture is cooled to0° C. MeOH (1 ml) and 1N—HCl (20 uL) are added carefully. After heatingfor 10 min at 80° C., the mixture is cooled and then basified with1N—NaOH to pH 10. The resulting mixture is extracted with DCM and thecombined extracts are washed with saturated brine. Drying over Na₂SO₄,and removal of the solvent in vacuo gave a yellow residue that ispurified by PTLC affording 19 mg of3-{4-[1-benzyl-ethyl-amino)-butyl]-isoquinolin-1-yl}-2,4-dimethyl-phenolas a white solid.

¹H NMR (CD₃Cl) 1:1 mixture of atropisomers 8.65 (d, 1H), 8.42 (br s,1H), 7.66 (t, 1H), 7.56 (d, 1H), 7.44 (t, 1H), 7.27-2.18 (m, 6H),6.89-6.86 (m, 1H), 6.71 (d, 1H), 4.48-4.43 (m, 1H), 3.68 (s, 2H),2.86-2.79 (m, 1H), 2.71-2.63 (m, 1H), 2.17-2.06 (m, 2H), 1.75 (2s, 3H),1.63 (2s, 3H), 1.30-1.21 (m, 2H), 1.06-1.00 (m, 3H), 0.93 (t, 3H).

LCMS (m/z):439.3 (MH)⁺

Example 6 Synthesis of3-[4-(1-ethoxy-butyl)-isoquinolin-1-yl]-2,4-dimethyl-phenol

Step 1. Preparation of1-[1-(3-Methoxy-2,6-dimethyl-phenyl)-isoquinolin-4-yl]-butan-1-ol

n-PrMgCl (4.7 ml of 2M solution in THF, 9.36 mmol) is added to asolution of1-(3-methoxy-2,6-dimethyl-phenyl)-isoquinoline-4-carbaldehyde (2.48 g,8.51 mmol) dropwise at −78 C and the mixture is allowed to warm to roomtemperature. After quenching with saturated NH₄Cl, the mixture isextracted with EtOAc. The combined extracts are washed with saturatedbrine, dried over Na₂SO₄, and concentrated. The residue ischromatographed on silica gel to give 1.9 g of1-[1-(3-methoxy-2,6-dimethyl-phenyl)-isoquinolin-4-yl]-butan-1-ol as awhite foam.

¹H NMR (CD₃Cl) 1:1 mixture of atropisomers 8.71 (s, 1H), 8.26 (dd, 1H),7.71 (t, 1H), 7.57 (d, 1H), 7.46 (t, 1H), 7.11 (d, 1H), 6.89 (d, 1H),5.43-5.39 (m, 1H), 3.87 (s, 3H), 2.21 (br s, 1H), 2.09-2.01 (m, 2H),1.80 (2s, 3H), 1.74 (2s, 3H), 1.70-1.46 (m 2H), 1.01 (t, 3H).

Step 2. Preparation of3-[4-(1-Bromo-butyl)-isoquinolin-1-yl]-2,4-dimethyl-phenol

A solution of1-[1-(3-methoxy-2,6-dimethyl-phenyl)-isoquinolin-4-yl]-butan-1-ol (117mg, 0.33 mmol) in DMF (2 ml) is added to a suspension of NaH (67 mg, 60%dispersion in mineral oil, 1.7 mmol) in DMF (4 ml) at 0° C. Afterstirring for 45 min, EtI (0.26 ml, 3.3 mmol) is added slowly. Afterstirring for 10 min at 0° C., the mixture is allowed to warm to roomtemperature and the stirring is continued for 1 h. Water is added andthe mixture is extracted with Et₂O. The combined extracts are washedwith saturated brine, dried over Na₂SO₄, and concentrated to give thedesired ether that is dissolved in DCM (12 ml). After cooling to −78°C., BBr₃ (3.2 ml of 1 M solution in DCM, 3.2 mmol) is added dropwise andthe mixture is allowed to warm to room temperature. After stirring for 1h, MeOH (3 ml) and 1N—HCl (50 uL) are added carefully. After heating for10 min at 80° C., the mixture is cooled and then basified with 1N—NaOHto pH 10. The resulting mixture is extracted with DCM and the combinedextracts are washed with saturated brine. Drying over Na₂SO₄ andconcentration are followed by flash chromatography to give 120 mg of3-[4-(1-bromo-butyl)-isoquinolin-1-yl]-2,4-dimethyl-phenol as a yellowfoam.

¹H NMR (CD₃Cl) 1:1 mixture of atropisomers 8.81 (s, 1H), 8.31 (d, 1H),7.82 (t, 1H), 7.64 (d, 1H), 7.52 (t, 1H), 7.20 (br s, 1H), 6.90 (d, 1H),6.73 (d, 1H), 5.71-5.66 (m, 1H), 2.71-2.58 (m, 1H), 2.50-2.38 (m, 1H),1.79 (2s, 3H), 1.68 (2s, 3H), 1.61-1.52 (m, 2H), 1.06 (t, 3H).

Step 3. Preparation of3-[4-(1-Ethoxy-butyl)-isoquinolin-1-yl]-2,4-dimethyl-phenol

A mixture of 3-[4-(1-bromo-butyl)-isoquinolin-1-yl]-2,4-dimethyl-phenol(110 mg, 0.29 mmol) and c-HCl (30 uL) in EtOH (12 ml) is heated atreflux for 18 h. After removal of solvent, the residue is diluted withDCM and neutralized with 1N—NaOH. The layers are separated and theaqueous layer is extracted with DCM. The combined organic layers aredried over Na₂SO₄ and concentrated. The residue is chromatographed onsilica gel to give 19 mg of3-[4-(1-Ethoxy-butyl)-isoquinolin-1-yl]-2,4-dimethyl-phenol along withthe unreacted starting material (80 mg).

¹H NMR (CD₃Cl) 8.62 (s, 1H), 8.39 (d, 1H), 7.71 (t, 1H), 7.62 (d, 1H),7.47 (t, 1H), 6.85 (d, 1H), 6.68 (d, 1H), 4.89 (t, 1H), 3.53-3.44 (m,2H), 2.12-2.05 (m, 1H), 1.99-1.89 (m, 1H), 1.78 (s, 3H), 1.65 (s, 3H),1.61-1.35 (m, 2H), 1.25 (t, 3H), 0.97 (t, 3H).

LCMS (m/z):350.2 (MH)⁺

Example 51 Additional 1-Aryl-4-Substituted Isoquinolines

The compounds shown in Table 1 are prepared according to the proceduresgiven in the above Schemes and further illustrated in the aboveExamples.

Unless otherwise specified all starting materials and reagents are ofstandard commercial grade, and are used without further purification, orare readily prepared from such materials by routine methods. Thoseskilled in the art of organic synthesis will recognize that startingmaterials and reaction conditions may be varied to achieve the desiredend product.

LC/MS data is provided in the tables, along with retention time inminutes and a number (1, 2 or 3) indicating the method used. The LC/MSmethods are as follows:

Method 1:

-   -   Analytical HPLC/MS instrumentation: Analyses are performed using        a Waters 600 series pump (Waters Corporation, Milford, Mass.), a        Waters 996 Diode Array Detector and a Gilson 215 auto-sampler        (Gilson Inc, Middleton, Wis.), Micromass® LCT time-of-flight        electrospray ionization mass analyzer. Data are acquired using        MassLynx™ 4.0 software, with OpenLynx Global Server™, OpenLynx™,        and AutoLynx™ processing.    -   Analytical HPLC conditions: 4.6×50 mm, Chromolith™ SpeedROD        RP-18e column (Merck KGaA, Darmstadt, Germany); UV 10        spectra/sec, 220-340 nm summed; flow rate 6.0 mL/min; injection        volume 1 μl;        -   Gradient conditions—mobile phase A is 95% water, 5% methanol            with 0.05% TFA; mobile phase B is 95% methanol, 5% water            with 0.025% TFA, and the gradient is 0-0.5 minutes 10-100%            B, hold at 100% B to 1.2 minutes, return to 10% B at 1.21            minutes inject-to-inject cycle time is 2.15 minutes.    -   Analytical MS conditions: capillary voltage 3.5 kV; cone voltage        30V; desolvation and source temperature are 350° C. and 120° C.,        respectively; mass range 181-750 with a scan time of 0.22        seconds and an inter scan delay of 0.05 minutes.        Method 2:    -   HPLC instrumentation: Analyses are performed using a Waters 600        series pump (Waters Corporation, Milford, Mass.), a Waters 996        Diode Array Detector and a Gilson 215 autosampler (Gilson Inc,        Middleton, Wis.). Data are acquired using MassLynx 4.0 software,        with OpenLynx processing.    -   HPLC conditions: 4.6×50 mm, Chromolith SpeedRod column (Merck        AEG); UV 5 spectra/sec, 220, 254 nm; flow rate 6.0 mL/min;        injection volume 1-10 μl;        -   Gradient conditions—Mobile phase A 95% Water, 5% Methanol            with 0.05% Formic acid; Mobile phase B 95% Methanol, 5%            Water with 0.025% Formic acid;

Gradient: Time(mins) % B 0 5 0.01 5 1.0 100 2 100 2.1 5

-   -   MS instrumentation: LC-MS experiments are performed using a        Waters ZMD II Mass Spectrometer.    -   MS conditions: Electrospray positive ionization; capillary        voltage 3.5 kV; cone voltage 30V; desolvation and source        temperature 250° C. and 100° C. respectively; mass range 120-800        with a scan time of 0.5 seconds and an inter scan delay of 0.1        mins.        Method 3:    -   HPLC instrumentation: Analyses are performed using a Waters 600        series pump (Waters Corp.), a Waters 996 Diode Array Detector        and a Gilson 215 autosampler (Gilson Inc.). Data are acquired        using MassLynx 4.0 software, with OpenLynx processing.    -   HPLC conditions: 4.6×50 mm, XTerra MS C18, 5 μm column (Waters        Corp.); UV 10 spectra/sec, 220, 254 nm; flow rate 4.0 mL/min;        injection volume 1-10 μl;        -   Gradient conditions—Mobile phase A 95% Water, 5% Methanol            with 0.05% Formic acid; Mobile phase B 95% Methanol, 5%            Water with 0.025% Formic acid;

Gradient: Time(mins) % B 0 5 0.01 5 2.0 100 3.50 100 3.51 5

-   -   MS instrumentation: LC-MS experiments are performed using a        Waters ZMD II Mass Spectrometer.    -   MS conditions: Electrospray positive ionization; capillary        voltage 3.5 kV; cone voltage 30V; desolvation and source        temperature 250° C. and 100° C. respectively; mass range 120-800        with a scan time of 0.5 seconds and an inter scan delay of 0.1        mins.

TABLE I LCMS Retention LCMS LCMS Time Mass M + H Cpd # Structure (min)(amu) (amu) 1

1.2 360.26 361.29 2

1.18 376.25 377.29 3

1.15 348.22 349.24 4

1.16 334.2 335.24 5

1.14 334.2 335.24 6

1.13 405.28 406.32 7

1.09 431.29 432.33 8

1.13 320.19 321.2 9

1.15 376.5 377.3 10

1.17 332.23 333.26 11

1.17 362.24 363.27 12

1.13 348.22 349.26 13

1.15 348.22 349.25 14

1.13 334.2 335.24 15

1.16 362.24 363.28 16

1.17 348.22 349.31 17

1.18 346.24 347.32 18

1.2 368.23 369.31 19

1.15 362.24 363.32 20

1.15 375.23 376.29 21

1.25 378.22 378.26 22

23

1.27 372.12 373.16 24

1.18 344.23 345.26 25

1.18 352.2 353.23 26

1.17 362.24 363.28 27

1.15 348.22 349.26 28

1.18 376.25 377.29 29

1.19 376.25 377.3 30

1.16 362.24 363.27 31

1.08 320.19 321.21 32

1.15 361.25 362.29 33

1.15 356.23 357.27 34

1.17 357.22 358.25 35

1.24 380.2 381.26 36

1.13 373.22 374.26 37

1.17 415.26 416.31 38

1.14 385.22 386.26 39

1.22 374.27 375.31 40

1.16 371.24 372.36 41

1.14 389.25 390.37 42

1.16 403.26 404.17 43

1.19 431.29 432.21 44

1.19 431.29 432.21 45

1.21 479.29 480.22 46

1.23 354.19 355.3 47

1.17 366.21 367.32 48

1.17 350.22 351.33 49

1.21 350.22 351.33 50

1.16 366.21 367.21 51

1.19 394.24 395.25 52

1.15 330.21 331.21 53

1.24 470.27 471.32 54

1.2 346.24 347.29 55

1.18 357.22 358.24 56

1.17 332.23 333.2 57

1.2 346.24 347.22 58

1.2 360.26 361.24 59

1.16 362.24 363.22 60

1.25 402.19 403.18 61

1.16 371.24 372.34 62

1.17 385.25 386.36 63

1.47 405.9 406.3 64

1.19 442.24 443.24 65

1.22 366.19 367.17 66

1.24 343.2 344.18 67

1.16 382.22 383.22 68

1.17 376.25 377.23 69

1.17 372.23 373.22 70

1.24 400.26 401.26 71

1.23 386.25 387.25 72

1.25 371.24 372.32 73

1.25 404.25 406.39 74

1.24 424.22 425.3 75

1.16 417.6 418.3 76

1.19 403.26 404.37 77

1.18 389.25 390.35

TABLE II 79

1.09 410.2 411.3 80

1.12 424.3 425.3 81

1.12 436.3 437.3 82

1.13 347.2 348.2 83

1.18 333.2 334.3 84

1.22 329.2 330.3 85

0.93 422.3 180.9 86

1.16 422.3 423.2 87

1.19 331.2 332.1 88

1.1 378.2 379.2 89

1.17 347.2 348.2 90

1.2 389.2 390.2 91

1.16 353.2 354.2 92

1.2 452.3 453.3 93

1.16 438.3 439.3 94

1.23 363.2 364.2 95

1.18 420.3 421.3 96

97

98

1.14 349.2 350.2 99

1.27 463.3 464.4 100

1.23 450.2 451.4 101

1.1 428.3 429.4 102

1.13 474.3 475.5 103

1.11 458.3 459.5 104

1.11 487.3 488.5 105

1.12 444.3 445.4

Example 52 Pharmaceutical Preparations of Oral and IntravenousAdministration

A. Tablets containing a C5a antagonist and an anti-arthritic agent thatis not a C5a receptor antagonist can be prepared as illustrated below:

Ingredient Amount C5a receptor antagonist 5 mg-500 mg C5areceptor-inactive therapeutic agent 1 mg-500 mg diluent, binder,disintigrant, lubricant, excipients q.s. 200-400 mg.B. Tablets containing a C5a receptor antagonist as the only activeingredient can be prepared as illustrated below:

Ingredient mg mg C5a receptor antagonist 10 50 MicrocrystallineCellulose 70.4 352 Granular Mannitol 15.1 75.5 Croscarmellose Sodium 3.015.0 Colloidal Silicon Dioxide 0.5 2.5 Magnesium Stearate (ImpalpablePowder) 1.0 5.0 Total (mg) 100 500C. Tablets containing a C5a receptor antagonist and a C5a receptorinactive agent may be prepared as follows:

Ingredient mg mg C5a receptor antagonist 10 25 C5a receptor inactivetherapeutic agent 10 25 Microcrystalline Cellulose 40 100 Modified foodcorn starch 1.05 4.25 Magnesium stearate 1.25 0.5D. Intravenous formulations containing a C5a receptor antagonist and aC5a receptor inactive agent may be prepared as follows:

Ingredient Amount C5a receptor antagonist 0.5-10 mg C5a receptorinactive therapeutic agent 0.5-10 mg Sodium Citrate 5-50 mg Citric Acid1-15 mg Sodium Chloride 1-8 mg Water for Injection to 1.0 literE. Oral suspensions containing a C5a receptor antagonist and a C5areceptor inactive agent may be prepared as follows:

Ingredient Amount per 5 mL dose C5a receptor antagonist 5-100 mg C5areceptor inactive 5-100 mg therapeutic agent Polyvinylpyrrolidone 150 mgPoly oxyethylene sorbitan 25 mg monolaurate Benzoic Acid 10 mg to 5 mLwith sorbitol solution (70%)

Example 53 Preparation of Radiolabeled Probes

Compounds provided herein are prepared as radiolabeled probes bycarrying out their synthesis using precursors comprising at least oneatom that is a radioisotope. The radioisotope is preferably at least oneof carbon (preferably ¹⁴C), hydrogen (preferably ³H), sulfur (preferably³⁵S), or iodine (preferably ¹²⁵I). Such radiolabeled probes areconveniently synthesized by a radioisotope supplier specializing incustom synthesis of radiolabeled probe compounds. Such suppliers includeAmersham Corporation, Arlington Heights, Ill.; Cambridge IsotopeLaboratories, Inc. Andover, Mass.; SRI International, Menlo Park,Calif.; Wizard Laboratories, West Sacramento, Calif.; ChemSynLaboratories, Lexena, Kans.; American Radiolabeled Chemicals, Inc., St.Louis, Mo.; and Moravek Biochemicals Inc., Brea, Calif.

Tritium labeled probe compounds are also conveniently preparedcatalytically via platinum-catalyzed exchange in tritiated acetic acid,acid-catalyzed exchange in tritiated trifluoroacetic acid, orheterogeneous-catalyzed exchange with tritium gas. Such preparations arealso conveniently carried out as a custom radiolabeling by any of thesuppliers listed in the preceding paragraph using a compound providedherein as substrate. In addition, certain precursors may be subjected totritium-halogen exchange with tritium gas, tritium gas reduction ofunsaturated bonds, or reduction using sodium borotritide, asappropriate.

Example 54 Receptor Autoradiography

Receptor autoradiography (receptor mapping) is carried out in vitro asdescribed by Kuhar in sections 8.1.1 to 8.1.9 of Current Protocols inPharmacology (1998) John Wiley & Sons, New York, using radiolabeledcompounds prepared as described herein.

Example 55 Assay for C5A Receptor Mediated Chemotaxis

This Example provides a standard assay of C5a receptor-mediatedchemotaxis.

Human promonocytic U937 cells (or purified human or non-humanneutrophils) are treated with dibutyryl cAMP for 48 hours prior toperforming the assay. Human neutrophils or those from another mammalianspecies are used directly after isolation. The cells are pelleted andresuspended in culture media containing 0.1% fetal bovine serum (FBS)and 10 μg/mL calcein AM (a fluorescent dye). This suspension is thenincubated at 37° C. for 30 minutes such that the cells take up thefluorescent dye. The suspension is then centrifuged briefly to pelletthe cells, which are then resuspended in culture media containing 0.1%FBS at a concentration of approximately 3×10⁶ cells/mL. Aliquots of thiscell suspension are transferred to clean test tubes, which containvehicle (1% DMSO in culture media containing 0.1% FBS) or varyingconcentrations of a compound of interest, and are incubated at roomtemperature for at least 30 minutes. The chemotaxis assay is performedin CHEMO TX 101-8, 96 well plates (Neuro Probe, Inc.; Gaithersburg,Md.). The bottom wells of the plate are filled with medium containing0-10 nM of C5a, preferably derived from the same species of mammal asare the neutrophils or other cells (e.g., human C5a for human U937cells). The top wells of the plate are filled with cell suspensions(compound- or vehicle-treated). The plate is then placed in a tissueculture incubator for 60 minutes. The top surface of the plate is washedwith PBS to remove excess cell suspension. The number of cells that havemigrated into the bottom well is then determined using a fluorescencereader. Chemotaxis index (the ratio of migrated cells to total number ofcells loaded) is then calculated for each compound concentration todetermine an EC₅₀ value.

As a control to ensure that cells retain chemotactic ability in thepresence of the compound of interest, the bottom wells of the plate maybe filled with varying concentrations chemo-attractants that do notmediate chemotaxis via the C5a receptor, such as zymosan-activated serum(ZAS), N-formylmethionyl-leucyl-phenylalanine (FMLP) or leukotriene B4(LTB4), rather than C5a, under which conditions compounds providedherein preferably do not detectably inhibit chemotaxis. Preferred C5areceptor modulators exhibit EC₅₀ values of less than 1 μM in the aboveassay for C5a mediated chemotaxis.

Example 56 Expression of A C5A Receptor

A human C5a receptor cDNA is obtained by PCR using 1) a forward primeradding a Kozak ribosome binding site and 2) a reverse primer that addsno additional sequence, and 3) an aliquot of a Stratagene Human FetalBrain cDNA library as template. The sequence of the resulting PCRproduct is described in PCT International Application WO 02/49993 as SEQID NO:1. The PCR product is subcloned into the cloning vector pCR-ScriptAMP (STRATAGENE, La Jolla, Calif.) at the Srf I site. It is then excisedusing the restriction enzymes EcoRI and NotI and subcloned in theappropriate orientation for expression into the baculoviral expressionvector pBacPAK 9 (CLONTECH, Palo Alto, Calif.) that has been digestedwith EcoRI and NotI.

Example 57 Baculoviral Preparations for C5A Expression

The human C5a (hC5a) receptor baculoviral expression vector isco-transfected along with BACULOGOLD DNA (BD PharMingen, San Diego,Calif.) into Sf9 cells. The SP cell culture supernatant is harvestedthree days post-transfection. The recombinant virus-containingsupernatant is serially diluted in Hink's TNM-FH insect medium (JRHBiosciences, Kansas City) supplemented Grace's salts and with 4.1 mML-Gln, 3.3 g/L LAH, 3.3 g/L ultrafiltered yeastolate and 10%heat-inactivated fetal bovine serum (hereinafter “insect medium”) andplaque assayed for recombinant plaques. After four days, recombinantplaques are selected and harvested into 1 mL of insect medium foramplification. Each 1 mL volume of recombinant baculovirus (at passage0) is used to infect a separate T25 flask containing 2×10⁶ Sf9 cells in5 mL of insect medium. After five days of incubation at 27° C.,supernatant medium is harvested from each of the T25 infections for useas passage 1 inoculum.

Two of seven recombinant baculoviral clones are then chosen for a secondround of amplification, using 1 mL of passage 1 stock to infect 1×10⁸cells in 100 mL of insect medium divided into 2 T175 flasks. Forty-eighthours post infection, passage 2 medium from each 100 mL prep isharvested and plaque assayed for titer. The cell pellets from the secondround of amplification are assayed by affinity binding as describedbelow to verify recombinant receptor expression. A third round ofamplification is then initiated using a multiplicity of infection of 0.1to infect a liter of Sf9 cells. Forty hours post-infection thesupernatant medium is harvested to yield passage 3 baculoviral stock.

The remaining cell pellet is assayed for affinity binding using theprotocol of DeMartino et al. (1994). J. Biol. Chem. 269(20):14446-14450(which is incorporated herein by reference for its teaching of bindingassays at page 14447), adapted as follows. Radioligand is 0.005-0.500 nM[¹²⁵I]C5a (human recombinant) (New England Nuclear Corp., Boston,Mass.); the hC5a receptor-expressing baculoviral cells are used insteadof 293 cells; the assay buffer contains 50 mM Hepes pH. 7.6, 1 mM CaCl₂,5 mM MgCl₂, 0.1% BSA, pH 7.4, 0.1 mM bacitracin, and 100 KIU/mLaprotinin; filtration is carried out using GF/C WHATMAN filters(presoaked in 1.0% polyethyeneimine for 2 hours prior to use); and thefilters are washed twice with 5 mL cold binding buffer without BSA,bacitracin, or aprotinin.

Titer of the passage 3 baculoviral stock is determined by plaque assayand a multiplicity of infection, incubation time course, binding assayexperiment is carried out to determine conditions for optimal receptorexpression.

A multiplicity of infection of 0.1 and a 72-hour incubation period werethe best infection parameters found for hC5a receptor expression in upto 1-liter Sf9 cell infection cultures.

Example 58 Baculoviral Infections

Log-phase Sf9 cells (INVITROGEN Corp., Carlsbad Calif.), are infectedwith one or more stocks of recombinant baculovirus followed by culturingin insect medium at 27° C. Infections are carried out either only withvirus directing the expression of the hC5a receptor or with this virusin combination with three G-protein subunit-expression virus stocks: 1)rat G□_(i2) G-protein-encoding virus stock (BIOSIGNAL #V5J008), 2)bovine b1 G-protein-encoding virus stock (BIOSIGNAL #V5H012), and 3)human g2 G-protein-encoding virus stock (BIOSIGNAL #V6B003), which maybe obtained from BIOSIGNAL Inc., Montreal.

The infections are conveniently carried out at a multiplicity ofinfection of 0.1:1.0:0.5:0.5. At 72 hours post-infection, a sample ofcell suspension is analyzed for viability by trypan blue dye exclusion,and the remaining Sf9 cells are harvested via centrifugation (3000rpm/10 minutes/4° C.).

Example 59 Purified Recombinant Insect Cell Membranes

Sf9 cell pellets are resuspended in homogenization buffer (10 mM HEPES,250 mM sucrose, 0.5 μg/mL leupeptin, 2 μg/mL Aprotinin, 200 μM PMSF, and2.5 mM EDTA, pH 7.4) and homogenized using a POLYTRON homogenizer(setting 5 for 30 seconds). The homogenate is centrifuged (536×g/10minutes/4° C.) to pellet the nuclei. The supernatant containing isolatedmembranes is decanted to a clean centrifuge tube, centrifuged(48,000×g/30 minutes, 4° C.) and the resulting pellet resuspended in 30mL homogenization buffer. This centrifugation and resuspension step isrepeated twice. The final pellet is resuspended in ice cold Dulbecco'sPBS containing 5 mM EDTA and stored in frozen aliquots at −80° C. untilneeded. The protein concentration of the resulting membrane preparation(hereinafter “P2 membranes”) is conveniently measured using a Bradfordprotein assay (Bio-Rad Laboratories, Hercules, Calif.). By this measure,a 1-liter culture of cells typically yields 100-150 mg of total membraneprotein.

Example 60 Radioligand Binding Assays

Purified P2 membranes, prepared by the method given above, areresuspended by Dounce homogenization (tight pestle) in binding buffer(50 mM Hepes pH. 7.6, 120 mM NaCl, 1 mM CaCl₂, 5 mM MgCl₂, 0.1% BSA, pH7.4, 0.1 mM bacitracin, 100 KIU/mL aprotinin).

For saturation binding analysis, membranes (5-50 μg) are added topolypropylene tubes containing 0.005-0.500 nM [¹²⁵I]C5a (human(recombinant), New England Nuclear Corp., Boston, Mass.) with a finalassay volume of 0.25 ml. Nonspecific binding is determined in thepresence of 300 nM hC5a (Sigma Chemical Co., St. Louis, Mo.) andaccounted for less than 10% of total binding. For evaluation of guaninenucleotide effects on receptor affinity, GTPγS is added to duplicatetubes at the final concentration of 50 μM.

For competition analysis, membranes (5-50 μg) are added to polypropylenetubes containing 0.030 nM [¹²⁵I]C5a (human). Non-radiolabeled displacersare added to separate assays at concentrations ranging from 10⁻¹⁰ M to10⁻⁵ M to yield a final volume of 0.250 mL. Nonspecific binding isdetermined in the presence of 300 nM hC5a (Sigma Chemical Co., St.Louis, Mo.) and accounted for less than 10% of total binding. Followinga 2-hour incubation at room temperature, the reaction is terminated byrapid vacuum filtration. Samples are filtered over presoaked (in 1.0%polyethyleneimine for 2 hours prior to use) GF/C WHATMAN filters andrinsed 2 times with 5 mL cold binding buffer without BSA, bacitracin, oraprotinin. Remaining bound radioactivity is quantified by gammacounting. K_(I) and Hill coefficient (“nH”) are determined by fittingthe Hill equation to the measured values with the aid of SIGMAPLOTsoftware.

Example 61 Agonist-Induced GTP Binding

Agonist-stimulated GTP-gamma³⁵S binding (“GTP binding”) activity can beused to identify agonist and antagonist compounds and to differentiateneutral antagonist compounds from those that possess inverse agonistactivity. This activity can also be used to detect partial agonismmediated by antagonist compounds. A compound being analyzed in thisassay is referred to herein as a “test compound.” Agonist-stimulated GTPbinding activity is measured as follows: Four independent baculoviralstocks (one directing the expression of the hC5a receptor and threedirecting the expression of each of the three subunits of aheterotrimeric G-protein) are used to infect a culture of Sf9 cells asdescribed above.

Agonist-stimulated GTP binding on purified membranes (prepared asdescribed above) is assessed using hC5a (Sigma Chemical Co., St. Louis,Mo., USA) as agonist in order to ascertain that thereceptor/G-protein-alpha-beta-gamma combination(s) yield a functionalresponse as measured by GTP binding.

P2 membranes are resuspended by Dounce homogenization (tight pestle) inGTP binding assay buffer (50 mM Tris pH 7.0, 120 mM NaCl, 2 mM MgCl₂, 2mM EGTA, 0.1% BSA, 0.1 mM bacitracin, 100KIU/mL aprotinin, 5 μM GDP) andadded to reaction tubes at a concentration of 30 μg protein/reactiontube. After adding increasing doses of the agonist hC5a atconcentrations ranging from 10⁻¹² M to 10⁻⁶ M, reactions are initiatedby the addition of 100 pM GTPgamma³⁵S with a final assay volume of 0.25ml. In competition experiments, non-radiolabeled test compounds (e.g.,compounds of Formula I) are added to separate assays at concentrationsranging from 10⁻¹⁰ M to 10⁻⁵M along with 10 nM hC5a to yield a finalvolume of 0.25 mL.

Neutral antagonists are those test compounds that reduce theC5a-stimulated GTP binding activity towards, but not below, baseline(the level of GTP bound by membranes in this assay in the absence ofadded C5a or other agonist and in the further absence of any testcompound).

In contrast, in the absence of added C5a, certain preferred compoundsreduce the GTP binding activity of the receptor-containing membranesbelow baseline, and are thus characterized as inverse agonists. If atest compound that displays antagonist activity does not reduce the GTPbinding activity below baseline in the absence of the C5a agonist, it ischaracterized as a neutral antagonist.

An antagonist test compound that elevates GTP binding activity abovebaseline in the absence of added hC5a in this assay is characterized ashaving partial agonist activity. Preferred antagonist compounds providedherein do not elevate GTP binding activity under such conditions morethan 10% above baseline, preferably not more than 5% above baseline, andmost preferably not more than 2% above baseline.

Following a 60-minute incubation at room temperature, the reactions areterminated by vacuum filtration over GF/C filters (pre-soaked in washbuffer, 0.1% BSA) followed by washing with ice-cold wash buffer (50 mMTris pH 7.0, 120 mM NaCl). The amount of receptor-bound (and therebymembrane-bound) GTPgamma³⁵S is determined by measuring the boundradioactivity, preferably by liquid scintillation spectrometry of thewashed filters. Non-specific binding is determined using 10 mM GTPgammaSand typically represents less than 5 percent of total binding. Data isexpressed as percent above basal (baseline). The results of these GTPbinding experiments is analyzed using SIGMAPLOT software (SPSS Inc.,Chicago, Ill.).

Example 62 Calcium Mobilization Assays

A. Response to C5a

U937 cells are grown in differentiation media (1 mM dibutyryl cAMP inRPMI 1640 medium containing 10% fetal bovine serum) for 48 hours at 37°C. then reseeded onto 96-well plates suitable for use in a FLIPR™ PlateReader (Molecular Devices Corp., Sunnyvale Calif.). Cells are grown anadditional 24 hours (to 70-90% confluence) before the assay. The cellsare then washed once with Krebs Ringer solution. FLUO-3 calciumsensitive dye (Molecular Probes, Inc. Eugene, Oreg.) is added to 10μg/mL and incubated with the cells in Krebs Ringer solution at roomtemperature for 1 to 2 hours. The 96 well plates are then washed toremove excess dye. Fluorescence responses, measured by excitation at 480nM and emission at 530 nM, are monitored upon the addition of human C5ato the cells to a final concentration of 0.01-30.0 nM, using the FLIPR™device (Molecular Devices). Differentiated U937 cells typically exhibitsignals of 5,000-50,000 Arbitrary Fluorescent Light Units in response toagonist stimulation.

B. Assays for Determination of ATP Responses

Differentiated U937 cells (prepared and tested as described above under“A. Response to C5a”) are stimulated by the addition of ATP (rather thanC5a) to a final concentration of 0.01 to 30 μM. This stimulationtypically triggers a signal of 1,000 to 12,000 arbitrary fluorescencelight units. Certain preferred compounds produce less than a 10%,preferably less than a 5%, and most preferably less than a 2% alterationof this calcium mobilization signal when this control assay is carriedout in the presence or absence of the compounds.

C. Assays for the Identification of Receptor Modulatory Agents:Antagonists and Agonists

Those of skill in the art will recognize that the calcium mobilizationassay described above may be readily adapted for identifying testcompounds as having agonist or antagonist activity at the human C5areceptor.

For example, in order to identify antagonist compounds, differentiatedU937 cells are washed and incubated with Fluo-3 dye as described above.One hour prior to measuring the fluorescence signal, a subset of thecells is incubated with a 1 μM concentration of at least one compound tobe tested. The fluorescence response upon the subsequent addition of 0.3nM (final concentration) human recombinant C5a is monitored using theFLIPR™ plate reader. Antagonist compounds elicit at least a 2-folddecrease in the fluorescence response relative to that measured in thepresence of human C5a alone. Preferred antagonist compounds elicit atleast a 5-fold, preferably at least a 10-fold, and more preferably atleast a 20-fold decrease in the fluorescence response relative to thatmeasured in the presence of human C5a alone. Agonist compounds elicit anincrease in fluorescence without the addition of C5a, which increasewill be at least partially blocked by a known C5a receptor antagonist.

Example 63 Assays to Evaluate Agonist Activity of Small Molecule C5AReceptor Antagonists

Certain preferred compounds of Formula I are C5a receptor antagoniststhat do not possess significant (e.g., greater than 5%) agonist activityin any of the C5a mediated functional assays discussed herein. Suchagonist activity can be evaluated, for example, in the assay of C5ainduced GTP binding given above, by measuring small molecule mediatedGTP binding in the absence of the natural agonist, C5a. Similarly, in acalcium mobilization assay such as the assay described above a smallmolecule compound can be directly assayed for the ability of thecompound to stimulate calcium levels in the absence of the naturalagonist, C5a. The preferred extent of C5a agonist activity exhibited bycertain compounds provided herein is less than 10%, more preferably lessthan 5% and most preferably less than 2% of the response elicited by thenatural agonist, C5a.

Example 64 MDCK Toxicity Assay

This Example illustrates the evaluation of compound toxicity using aMadin Darby canine kidney (MDCK) cell cytotoxicity assay.

1 μL of test compound is added to each well of a clear bottom 96-wellplate (PACKARD, Meriden, Conn.) to give final concentration of compoundin the assay of 10 micromolar, 100 micromolar or 200 micromolar. Solventwithout test compound is added to control wells.

MDCK cells, ATCC no. CCL-34 (American Type Culture Collection, Manassas,Va.), are maintained in sterile conditions following the instructions inthe ATCC production information sheet. Confluent MDCK cells aretrypsinized, harvested, and diluted to a concentration of 0.1×10⁶cells/ml with warm (37° C.) medium (VITACELL Minimum Essential MediumEagle, ATCC catalog #30-2003). 100 μL of diluted cells is added to eachwell, except for five standard curve control wells that contain 100 μLof warm medium without cells. The plate is then incubated at 37° C.under 95% O₂, 5% CO₂ for 2 hours with constant shaking. Afterincubation, 50 μL of mammalian cell lysis solution” (available as acomponent of the PACKARD (Meriden, Conn.) ATP-LITE-M Luminescent ATPdetection kit) is added per well, the wells are covered with PACKARDTOPSEAL stickers, and plates are shaken at approximately 700 rpm on asuitable shaker for 2 minutes.

Compounds causing toxicity will decrease ATP production, relative tountreated cells. The PACKARD ATP-LITE-M Luminescent ATP detection kit,product no. 6016941, is generally used according to the manufacturer'sinstructions to measure ATP production in treated and untreated MDCKcells. PACKARD ATP LITE-M reagents are allowed to equilibrate to roomtemperature. Once equilibrated, the lyophilized substrate solution isreconstituted in 5.5 mL of substrate buffer solution (from kit).Lyophilized ATP standard solution is reconstituted in deionized water togive a 10 mM stock. For the five control wells, 10 μL of seriallydiluted PACKARD standard is added to each of the standard curve controlwells to yield a final concentration in each subsequent well of 200 nM,100 nM, 50 nM, 25 nM and 12.5 nM. PACKARD substrate solution (50 μL) isadded to all wells, which are then covered, and the plates are shaken atapproximately 700 rpm on a suitable shaker for 2 minutes. A whitePACKARD sticker is attached to the bottom of each plate and samples aredark adapted by wrapping plates in foil and placing in the dark for 10minutes. Luminescence is then measured at 22° C. using a luminescencecounter (e.g., PACKARD TOPCOUNT Microplate Scintillation andLuminescence Counter or TECAN SPECTRAFLUOR PLUS), and ATP levelscalculated from the standard curve. ATP levels in cells treated withtest compound(s) are compared to the levels determined for untreatedcells. Cells treated with 10 μM of a preferred test compound exhibit ATPlevels that are at least 80%, preferably at least 90%, of the untreatedcells. When a 100 μM concentration of the test compound is used, cellstreated with preferred test compounds exhibit ATP levels that are atleast 50%, preferably at least 80%, of the ATP levels detected inuntreated cells.

1. A compound according to Formula I

or a pharmaceutically acceptable salt thereof, wherein: R₁ is selectedfrom hydrogen, cyano, amino, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted cycloalkyl, optionally substituted cycloalkenyl, optionallysubstituted haloalkyl, optionally substituted haloalkoxy, optionallysubstituted alkoxy, optionally substituted cycloalkoxy, or optionallysubstituted (cycloalkyl)alkoxy; R₂ is selected from the group consistingof —NR₄R₅, —(CR_(A)R_(B))OR₄, —CR_(A)R_(B)NR₄R₅, —C(R_(A′))═CR_(A)R_(B),and —CR_(A)R_(B)Q; R₃ represents between 0 and 4 substituents, each ofwhich is independently selected from halogen, hydroxy, amino, cyano,optionally substituted alkyl, optionally substituted haloalkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cycloalkyl, optionally substituted alkoxy,optionally substituted haloalkoxy, optionally substituted hydroxyalkyl,optionally substituted alkoxyalkyl, optionally substituted mono- anddi-alkylamino, optionally substituted aminoalkyl,-E-(CR_(C)R_(D))_(m)—Z, and -E-(CR_(C)R_(D))_(m)—XR_(A); R₄ is: (i)C₂-C₈alkyl, C₂-C₈alkenyl, C₂-C₈alkynyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl,mono- or di-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl, arylC₀-C₄alkyl, or (heteroaryl)C₀₋₄alkyl,each of which is substituted with from 0 to 4 substituents independentlychosen from R_(x), C₂-C₄alkanoyl, mono- anddi-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl and XR_(y); or (ii) joined to R₅ to form,with the nitrogen to which R₄ and R₅ are bound, a heterocycle havingfrom 1 to 3 rings, 5 to 7 ring members in each ring, wherein theheterocycle is substituted with from 0 to 4 substituents independentlychosen from R_(x), oxo and W—Z; R₅ is: (i) hydrogen; (ii) C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, (C₃-C₇-carbocycle)C₀-C₄alkyl, each of whichis substituted with from 0 to 3 substituents independently chosen fromhalogen, hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy, methylamino,dimethylamino, trifluoromethyl and trifluoromethoxy; or (iii) joined toR₄ to form an optionally substituted heterocycle; Ar is mono-, di-, ortri-substituted phenyl, optionally substituted naphthyl, or optionallysubstituted heteroaryl selected from pyridyl, furanyl, indolyl,pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, thienyl,thiazolyl, triazolyl, isoxazolyl, pyrrolyl, pyrazolyl,5,6,7,8-tetrahydroisoquinoline, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl,indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl,isoindazolyl, isoindolinyl, isoindolyl, isothiazolyl, isoxazolyl, andindolin-2-one,  wherein Ar is optionally substituted heteroaryl when R₂is —NR₄R₅; R_(A), R_(A′), and R_(B), which may be the same or different,are independently selected at each occurrence from: (i) hydrogen andhydroxy, and (ii) alkyl groups, cycloalkyl groups, and (cycloalkyl)alkylgroups, each of which is optionally substituted with one or moresubstituent(s) independently selected from oxo, hydroxy, halogen, cyano,amino, C₁₋₆alkoxy, —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)(C₁₋₆alkyl),—NHC(═O)(C₁₋₆alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆alkyl),—NHS(O)_(n)(C₁₋₆alkyl), —S(O)_(n)(C₁₋₆ alkyl), —S(O)_(n)NH(C₁₋₆alkyl),—S(O)_(n)N(C₁₋₆ alkyl)(C₁₋₆alkyl), and Z; E is a single covalent bond,oxygen, or NR_(A); R_(x) is independently chosen at each occurrence fromhalogen, hydroxy, amino, cyano, nitro, —COOH, —C(═O)NH₂,C₁-C₆alkoxycarbonyl, mono- and di-(C₁₋₆alkyl)aminocarbonyl, C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, mono- and di-(C₁-C₆alkyl)amino, C₁-C₆alkoxy,C₁-C₂hydroxyalkyl, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,(C₃-C₇cycloalkyl)C₀-C₄alkyl, and —S(O_(n))C₁-C₆alkyl; X is independentlyselected at each occurrence from the group consisting of —CH₂—,—CHR_(B)—, —O—, —C(═O)—, —C(═O)O—, —S(O)_(n)—, —NH—, —NR_(B)—,—C(═O)NH—, —C(═O)NR_(B)—, —S(O)_(n)NH—, —S(O)_(n)NR_(B)—, —NHC(═O)—,—NR_(B)C(═O)—, —NHS(O)_(n)—, and —NR_(B)S(O)_(n)—; R_(y) is: (i)hydrogen; or (ii) C₁-C₁₀alkyl, C₂-C₁₀alkenyl, C₂-C₁₀alkynyl,C₃-C₁₀-carbocycleC₀-C₄alkyl or (3- to 10-memberedheterocycle)C₀-C₄alkyl, each of which is substituted with from 0 to 6substituents independently selected from R_(x), oxo, —NH(C₁-C₆alkanoyl),—N(C₁-C₆alkyl)C₁-C₆alkanoyl, —NHS(O_(n))C₁-C₆alkyl,—N(S(O_(n))C₁-C₆alkyl)₂, —S(O_(n))NHC₁-C₆alkyl and—S(O_(n))N(C₁-C₆alkyl)₂; Y and Z are independently selected at eachoccurrence from 3- to 7-membered carbocyclic or heterocyclic groupswhich are saturated, unsaturated, or aromatic, which are optionallysubstituted with one or more substituents independently selected fromhalogen, oxo, hydroxy, amino, cyano, C₁₋₄alkyl, —O(C₁₋₄alkyl),—NH(C₁₋₄alkyl), —N(C₁₋₄alkyl)(C₁₋₄alkyl), and —S(O)_(n)(alkyl); Q is anoptionally substituted carbocyclic or optionally substitutedheterocyclic group which are saturated, unsaturated or aromatic andcomprises between 3 and 18 ring atoms arranged in 1, 2, or 3 rings whichare fused, spiro or coupled by a bond; m is independently selected ateach occurrence from integers ranging from 0 to 8; and n is an integerindependently selected at each occurrence from 0, 1, and
 2. 2. Acompound according to Formula II:

or a pharmaceutically acceptable salt thereof, wherein: Ar issubstituted phenyl, optionally substituted naphthyl, or optionallysubstituted heteroaryl selected from pyridyl, furanyl, indolyl,pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, thienyl,thiazolyl, triazolyl, isoxazolyl, pyrrolyl, pyrazolyl,5,6,7,8-tetrahydroisoquinoline, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl,indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl,isoindazolyl, isoindolinyl, isoindolyl, isothiazolyl, isoxazolyl, andindolin-2-one; A is OR₄, NR₄R₅, or CR₄(XR_(y))₂; R₁ is chosen from: (i)hydrogen, amino, and cyano; and (ii) C₁-C₆alkyl, C₂-C₆alkenyl,C₂-C₆alkynyl, C₁-C₆alkoxy, C₁-C₄haloalkyl, C₁-C₄haloalkoxy, mono- anddi-(C₁-C₆alkyl)amino, (C₃-C₇cycloalkyl)C₀-C₄alkyl, and—S(O_(n))C₁-C₄alkyl, each of which is substituted with from 0 to 4substituents independently chosen from R_(x); R₃ represents between 0and 4 substituents, each of which is independently selected fromhydrogen, halogen, hydroxy, amino, cyano, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted cycloalkyl, optionally substituted alkoxy,optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl,optionally substituted mono- and di-alkylamino, optionally substitutedaminoalkyl, optionally substituted cycloalkyloxy, optionally substitutedaryl, optionally substituted arylalkyl, optionally substituted aryloxy,optionally substituted arylalkyloxy, optionally substituted heterocycle,optionally substituted heterocycle-oxy, -E-(CR_(C)R_(D))_(m)—Z,-E-(CR_(C)R_(D))_(m)—XR_(A); R₄ is: (i) C₂-C₈alkyl, C₂-C₈alkenyl,C₂-C₈alkynyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, mono- ordi-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl, arylC₀-C₄alkyl, or heteroarylC₀₋₄alkyl,each of which is substituted with from 0 to 4 substituents independentlychosen from R_(x), C₂-C₄alkanoyl, mono- anddi-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl and XR_(y); or (ii) joined to R₅ to form,with the nitrogen to which R₄ and R₅ are bound, a heterocycle havingfrom 1 to 3 rings, 5 to 7 ring members in each ring, wherein theheterocycle is substituted with from 0 to 4 substituents independentlychosen from R_(x), oxo and W—Z; R₅ is: (i) hydrogen; (ii) C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, (C₃-C₇-carbocycle)C₀-C₄alkyl, each of whichis substituted with from 0 to 3 substituents independently chosen fromhalogen, hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy, methylamino,dimethylamino, trifluoromethyl and trifluoromethoxy; or (iii) joined toR₄ to form an optionally substituted heterocycle; R₈ and R₉ areindependently selected from: (i) hydrogen, halogen, hydroxy, C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy, C₁-C₆alkylamino orC₃-C₇cycloalkyl C₀-C₄alkyl; E is a single covalent bond, oxygen, orNR_(A); X is a single covalent bond, —CR_(A)R_(B)—, —O—, —C(═O)—,—C(═O)O—, —S(O)_(n)— or —NR_(B)—; and R_(y) is: (i) hydrogen; or (ii)C₁-C₁₀alkyl, C₂-C₁₀alkenyl, C₂-C₁₀alkynyl, C₃-C₁₀-carbocycleC₀-C₄alkylor (3- to 10-membered heterocycle)C₀-C₄alkyl, each of which issubstituted with from 0 to 6 substituents independently selected fromR_(x), oxo, —NH(C₁-C₆alkanoyl), —N(C₁-C₆alkyl)C₁-C₆alkanoyl,—NHS(O_(n))C₁-C₆alkyl, —N(S(O_(n))C₁-C₆alkyl)₂, —S(O_(n))NHC₁-C₆alkyland —S(O_(n))N(C₁-C₆alkyl)₂; W is a single covalent bond, —CR_(A)R_(B)—,—NR_(B)— or —O—; Z is independently selected at each occurrence from 3-to 7-membered carbocycles and heterocycles, each of which is substitutedwith from 0 to 4 substituents independently selected from halogen, oxo,—COOH, hydroxy, amino, cyano, C₁-C₆alkyl, C₁-C₆alkoxy, C₁-C₆haloalkyl,C₁-C₆haloalkoxy, mono- and di-(C₁-C₆alkyl)amino and —S(O_(n))C₁-C₆alkyl; and R_(A) and R_(B) are independently selected at eachoccurrence from: (i) hydrogen; and (ii) C₁-C₁₀alkyl, C₂-C₁₀alkenyl,C₂-C₁₀alkynyl, saturated or partially saturated(C₃-C₁₀carbocycle)C₀-C₄alkyl and saturated or partially saturated (3- to10-membered heterocycle)C₀-C₄alkyl, each of which is substituted withfrom 0 to 6 substituents independently selected from oxo, hydroxy,halogen, cyano, amino, C₁-C₆alkoxy, mono- and di-(C₁-C₄alkyl)amino,—COOH, —C(═O)NH₂, —NHC(═O)(C₁-C₆alkyl), —N(C₁-C₆alkyl)C(═O)(C₁-C₆alkyl),—NHS(O_(n))C₁-C₆alkyl, SO₃H, —S(O_(n))C₁-C₆alkyl, —S(O_(n))NHC₁-C₆alkyl,—S(O_(n))N(C₁-C₆alkyl)C₁-C₆alkyl and Z; R_(C) and R_(D) areindependently selected from R_(A), hydroxy, C₁₋₆alkoxy, and oxo; R_(x)is independently chosen at each occurrence from halogen, hydroxy, amino,cyano, nitro, —COOH, —C(═O)NH₂, C₁-C₆alkoxycarbonyl, mono- anddi-(C₁₋₆alkyl)aminocarbonyl, C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,mono- and di-(C₁-C₆alkyl)amino, C₁-C₆alkoxy, C₁-C₂hydroxyalkyl,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, (C₃-C₇cycloalkyl)C₀-C₄alkyl, and—S(O_(n))C₁-C₆alkyl; m is an integer independently selected at eachoccurrence from 0-8; and n is an integer independently selected at eachoccurrence from 0, 1 and
 2. 3. A compound or pharmaceutically acceptablesalt thereof according to claim 2, wherein R₄ is: (i) C₂-C₈alkyl,C₂-C₈alkenyl, C₂-C₈alkynyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, mono- ordi-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl, phenylC₀-C₄alkyl, pyridylC₀-C₄alkyl,pyrimidinylC₀-C₄alkyl, thienylC₀-C₄alkyl, imidazolylC₀-C₄alkyl,pyrrolylC₀-C₄alkyl, pyrazolylC₀-C₄alkyl, benzoisothiazolyl ortetrahydronapthyl, each of which is substituted with from 0 to 4substituents independently chosen from R_(x), C₂-C₄alkanoyl, mono- anddi-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl and XR_(y); or (ii) joined to R₅ to form,with the nitrogen to which R₄ and R₅ are bound, a heterocycle havingfrom 1 to 3 rings, 5 to 7 ring members in each ring, wherein theheterocycle is substituted with from 0 to 4 substituents independentlychosen from R_(x), oxo and W—Z; and R₅ is: (i) hydrogen; (ii)C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, (C₃-C₇carbocycle)C₀-C₄alkyl,each of which is substituted with from 0 to 3 substituents independentlychosen from halogen, hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy,methylamino, dimethylamino, trifluoromethyl and trifluoromethoxy; or(iii) joined to R₄ to form an optionally substituted heterocycle.
 4. Acompound or pharmaceutically acceptable salt thereof according to claim2, wherein A is NR₄R₅.
 5. A compound or pharmaceutically acceptable saltthereof according to claim 4, wherein: R₄ is chosen from(C₃-C₇cycloalkyl)C₀-C₄alkyl, phenylC₀-C₄alkyl, pyridylC₀-C₄alkyl,pyrimidinylC₀-C₄alkyl, thienylC₀-C₄alkyl, imidazolylC₀-C₄alkyl,pyrrolylC₀-C₄alkyl, pyrazolylC₀-C₄alkyl, indolylC₀-C₄alkyl,indazolylC₀-C₄alkyl, benzocycloalkenylC₀-C₄alkyl,decahydronaphthylC₀-C₄alkyl, benzoisothiazolylC₀-C₄alkyl,tetrahydroquinolinylC₀-C₄alkyl and tetrahydronaphthylC₀-C₄alkyl, each ofwhich is substituted with from 0 to 4 groups independently chosen fromR_(x), mono- and di-C₁-C₄alkylamino(C₁-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl, C₂-C₄alkanoyl and C₂-C₄alkanoyloxy; and R₅is C₁-C₆alkyl, C₂-C₆alkenyl or (C₃-C₇carbocycle)C₀-C₄alkyl.
 6. Acompound or pharmaceutically acceptable salt thereof according to claim4, wherein R₄ and R₅ are joined to form a saturated or partiallysaturated heterocycle containing 1 or 2 fused or spiro rings; whereinthe heterocycle is substituted with from 0 to 4 substituentsindependently chosen from halogen, hydroxy, amino, cyano, —COOH,—CH₂COOH, C₁₋₆alkoxycarbonyl, —CH₂CO₂—C₁₋₆alkyl, —C(═O)NH₂, C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, mono- and di-(C₁-C₆alkyl)amino, C₁-C₆alkoxy,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, (C₃-C₇cycloalkyl)C₀-C₄alkyl,—S(O_(n))C₁-C₆alkyl, SO₃H, and phenyl.
 7. A compound or pharmaceuticallyacceptable salt thereof according to claim 4, wherein the compound isaccording to Formula III:

wherein: R₃ and R_(3a) are independently selected from hydrogen,C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, C₁₋₆haloalkoxy, COOH, CONH₂,SO₂NH₂, hydroxy, halogen, or amino; R₁₃ represents from 0 to 3substituents independently chosen from: (i) R_(x); and (ii) phenyl andpyridyl, each of which is substituted with from 0 to 4 substituentsindependently chosen from halogen, hydroxy, amino, cyano, C₁-C₄alkyl,C₁-C₄alkoxy, (C₃-C₇cycloalkyl)C₀-C₄alkyl, C₁-C₂haloalkyl,C₁-C₂haloalkoxy, and mono- and di-(C₁-C₄alkyl)amino; and G is CH₂,sulfur, oxygen or NR_(E); wherein R_(E) is: (i) hydrogen; or (ii)C₁-C₆alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, phenyl or a 5- or 6-memberedheteroaryl ring, each of which is substituted with from 0 to 3substituents independently chosen from R_(x).
 8. A compound orpharmaceutically acceptable salt thereof according to claim 4, whereinthe compound has the formula:

wherein: R₃ and R_(3a) are independently selected from the groupconsisting of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,C₁₋₆haloalkoxy, COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino; R₁₀and R₁₁ are independently chosen from hydrogen, C₁-C₆alkyl,C₁-C₂haloalkyl and C₃-C₇cycloalkyl(C₀-C₂alkyl); and R₁₂ represents from0 to 3 substituents independently chosen from R_(x), mono- anddi-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- anddi-(C₁-C₄alkyl)amino(C₁-C₄alkoxy) and YZ; or two adjacent R₁₂ groups arejoined to form a fused 5- to 7-membered carbocyclic or heterocyclicring.
 9. A compound or pharmaceutically acceptable salt thereofaccording to claim 4, wherein the compound has the formula:

wherein: R₃ and R_(3a) are independently selected from the groupconsisting of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,C₁₋₆haloalkoxy, COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino; R₁₂and R₁₃ independently represent from 0 to 3 substituents independentlychosen from R_(x); R₁₄ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl,C₂-C₆alkynyl, C₁-C₂haloalkyl or (C₃-C₇cycloalkyl)C₀-C₂alkyl, COOH,CONH₂, CH₂COOH, CH₂CONH₂, C₁₋₆alkoxycarbonyl, CH₂CO₂—C₁₋₆alkyl, or SO₃H;and x is 0, 1 or
 2. 10. A compound or pharmaceutically acceptable saltthereof according to claim 4, wherein the compound has the formula:

wherein: R₃ and R_(3a) are independently selected from the groupconsisting of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,C₁₋₆haloalkoxy, COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino; R₁₂and R₁₃ represent from 0 to 3 substituents independently chosen fromR_(x); G is CH₂, NH, sulfur or oxygen; G₃ is N, CH, or CR_(x) and x is0, 1 or
 2. 11. A compound or pharmaceutically acceptable salt thereofaccording to claim 4, wherein the compound has the formula:

wherein R₃ and R_(3a) are independently selected from the groupconsisting of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,C₁₋₆haloalkoxy, COOH, CONH₂, SO₂NH₂, hydroxy, halogen, and amino; R₁₂and R₁₃ independently represent from 0 to 3 substituents independentlychosen from R_(x); G is CH₂, NH or oxygen; and x is 0, 1 or
 2. 12. Acompound or pharmaceutically acceptable salt thereof according to claim2, wherein: A is OR₄; and R₄ is C₂-C₆alkyl, C₂-C₆alkenyl,phenylC₀-C₄alkyl, naphthylC₀-C₄alkyl, pyridylC₀-C₄alkyl,pyrimidinylC₀-C₄alkyl, thienylC₀-C₄alkyl, imidazolylC₀-C₄alkyl orpyrrolylC₀-C₄alkyl, each of which is substituted with from 0 to 4substituents independently chosen from R_(x), mono- anddi-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl and C₂-C₄alkanoyl.
 13. A compound orpharmaceutically acceptable salt thereof according to claim 1, whereinR₂ is —NR₄R₅; and Ar is heteroaryl.
 14. A compound according to FormulaIX:

or a pharmaceutically acceptable salt thereof, wherein: R₁ is selectedfrom the group consisting of hydrogen, halogen, cyano, amino, optionallysubstituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted cycloalkyl, optionallysubstituted cycloalkenyl, optionally substituted haloalkyl, optionallysubstituted haloalkoxy, optionally substituted alkoxy, optionallysubstituted cycloalkoxy, optionally substituted (cycloalkyl)alkoxy, andoptionally substituted heterocycloalkyl; R₃ represents between 0 and 4substituents, each of which is independently selected from optionallysubstituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted alkoxy, optionallysubstituted hydroxyalkyl, optionally substituted aminoalkyl, optionallysubstituted mono- and di-alkylamino, —O—(CR_(A)R_(B))_(m)—XR_(A),—O—(CR_(A)R_(B))_(m)—Y, —N(R_(B))—(CR_(A)R_(B))_(m)—XR_(A),—N(R_(B))—(CR_(A)R_(B))_(m)—Y; R₄ is: (i) C₂-C₈alkyl, C₂-C₈alkenyl,C₂-C₈alkynyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, mono- ordi-(C₁-C₄alkylamino)C₂-C₄alkyl, (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl, arylC₀-C₄alkyl, or (heterocycle)C₀₋₄alkyl,each of which is substituted with from 0 to 4 substituents independentlychosen from R_(x), C₂-C₄alkanoyl, mono- anddi-(C₁-C₄alkyl)amino(C₁-C₄alkyl), mono- anddi-C₁-C₄alkylamino(C₁-C₄alkoxy), (3- to 7-memberedheterocycloalkyl)C₀-C₄alkyl and XR_(y); or (ii) joined to R₅ to form,with the nitrogen to which R₄ and R₅ are bound, a heterocycle havingfrom 1 to 3 rings, 5 to 7 ring members in each ring, wherein theheterocycle is substituted with from 0 to 4 substituents independentlychosen from R_(x), oxo and W—Z; R₅ is: (i) hydrogen; (ii) C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, (C₃-C₇carbocycle)C₀-C₄alkyl, each of whichis substituted with from 0 to 3 substituents independently chosen fromhalogen, hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy, methylamino,dimethylamino, trifluoromethyl and trifluoromethoxy; or (iii) joined toR₄ to form an optionally substituted heterocycle; Ar is optionallysubstituted heteroaryl, selected from pyridyl, furanyl, indolyl,pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, thienyl,thiazolyl, triazolyl, isoxazolyl, pyrrolyl, pyrazolyl,5,6,7,8-tetrahydroisoquinoline, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl,indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl,isoindazolyl, isoindolinyl, isoindolyl, isothiazolyl, isoxazolyl, andindolin-2-one; R_(A) and R_(B), which may be the same or different, areindependently selected at each occurrence from: hydrogen, hydroxy, andstraight or branched alkyl groups, cycloalkyl groups, (cycloalkyl)alkylgroups and are optionally further substituted with one or moresubstituent(s) independently selected from oxo, hydroxy, halogen, cyano,amino, C₁₋₆alkoxy, —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)(C₁₋₆alkyl),—NHC(═O)(C₁₋₆alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆alkyl),—NHS(O)_(n)(C₁₋₆alkyl), —S(O)_(n)(C₁₋₆ alkyl), —S(O)_(n)NH(C₁₋₆alkyl),—S(O)_(n)N(C₁₋₆ alkyl)(C₁₋₆alkyl), and Z; X is independently selected ateach occurrence from the group consisting of —CH₂—, —CHR_(B)—, —O—,—C(═O)—, —C(═O)O—, —S(O)_(n)—, —NH—, —NR_(B)—, —C(═O)NH—, —C(═O)NR_(B)—,—S(O)_(n)NH—, —S(O)_(n)NR_(B)—, —NHC(═O)—, —NR_(B)C(═O)—, —NHS(O)_(n)—,and —NR_(B)S(O)_(n)—; Y and Z are independently selected at eachoccurrence from 3- to 7-membered carbocyclic or heterocyclic groupswhich are saturated, unsaturated, or aromatic, which are optionallysubstituted with one or more substituents independently selected fromhalogen, oxo, hydroxy, amino, cyano, C₁₋₄alkyl, —O(C₁₋₄alkyl),—NH(C₁₋₄alkyl), ₄alkyl), and —S(O)_(n)(alkyl), m is an integerindependently selected at each occurrence from integers in range of 0-8;and n is an integer independently selected at each occurrence from 0, 1,and
 2. 15. A pharmaceutical composition comprising at least one compoundor salt thereof according to claim 2, in combination with aphysiologically acceptable carrier or excipient.
 16. A packagedpharmaceutical preparation, comprising: (a) a pharmaceutical compositionaccording to claim 2 in a container; and (b) instructions for using thecomposition to treat a patient suffering from inflammation.